DIARYL SUBSTITUTED 6,5-FUSED RING COMPOUNDS AS C5aR INHIBITORS

ABSTRACT

or pharmaceutically acceptable salts thereof that are modulators of the C5a receptor. Also provided are pharmaceutical compositions and methods of use including the treatment of diseases or disorders involving pathologic activation from C5a and non-pharmaceutical applications.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is an application claiming benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62/609,834 filed Dec. 22,2017, which is herein incorporatd by reference in its entirety.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

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BACKGROUND OF THE INVENTION

The complement system plays a central role in the clearance of immunecomplexes and in immune responses to infectious agents, foreignantigens, virus infected cells and tumor cells. Inappropriate orexcessive activation of the complement system can lead to harmful, andeven potentially life-threatening consequences due to severeinflammation and resulting tissue destruction. These consequences areclinically manifested in various disorders including septic shock;myocardial, as well as, intestinal ischemia/reperfusion injury; graftrejection; organ failure; nephritis; pathological inflammation; andautoimmune diseases.

The complement system is composed of a group of proteins that arenormally present in the serum in an inactive state. Activation of thecomplement system encompasses mainly three distinct pathways, i.e., theclassical, the alternative, and the lectin pathway (V. M. Holers, InClinical Immunology: Principles and Practice, ed. R. R. Rich, MosbyPress; 1996, 363-391): 1) The classical pathway is acalcium/magnesium-dependent cascade, which is normally activated by theformation of antigen-antibody complexes. It can also be activated in anantibody-independent manner by the binding of C-reactive protein,complexed with ligand, and by many pathogens including gram-negativebacteria. 2) The alternative pathway is a magnesium-dependent cascadewhich is activated by deposition and activation of C₃ on certainsusceptible surfaces (e.g. cell wall polysaccharides of yeast andbacteria, and certain biopolymer materials). 3) The lectin pathwayinvolves the initial binding of mannose-binding lectin and thesubsequent activation of C₂ and C₄, which are common to the classicalpathway (Matsushita, M. et al., J. Exp. Med. 176: 1497-1502 (1992);Suankratay, C. et al., J. Immunol. 160: 3006-3013 (1998)).

The activation of the complement pathway generates biologically activefragments of complement proteins, e.g. C₃a, C₄a and C5a anaphylatoxinsand C₅b-9 membrane attack complexes (MAC), all which mediateinflammatory responses by affecting leukocyte chemotaxis; activatingmacrophages, neutrophils, platelets, mast cells and endothelial cells;and increasing vascular permeability, cytolysis and tissue injury.

Complement C5a is one of the most potent proinflammatory mediators ofthe complement system. (The anaphylactic C5a peptide is 100 times morepotent, on a molar basis, in eliciting inflammatory responses than C₃a.)C5a is the activated form of C₅ (190 kD, molecular weight). C5a ispresent in human serum at approximately 80 μg/ml (Kohler, P. F. et al.,J. Immunol. 99: 1211-1216 (1967)). It is composed of two polypeptidechains, α and β, with approximate molecular weights of 115 kD and 75 kD,respectively (Tack, B. F. et al., Biochemistry 18: 1490-1497 (1979)).Biosynthesized as a single-chain promolecule, C₅ is enzymaticallycleaved into a two-chain structure during processing and secretion.After cleavage, the two chains are held together by at least onedisulphide bond as well as noncovalent interactions (Ooi, Y. M. et al.,J. Immunol. 124: 2494-2498(1980)).

C₅ is cleaved into the C5a and C₅b fragments during activation of thecomplement pathways. The convertase enzymes responsible for C₅activation are multi-subunit complexes of C₄b, C₂a, and C₃b for theclassical pathway and of (C₃b)2, Bb, and P for the alternative pathway(Goldlust, M. B. et al., J. Immunol. 113: 998-1007 (1974); Schreiber, R.D. et al, Proc. Natl. Acad. Sci. 75: 3948-3952 (1978)). C₅ is activatedby cleavage at position 74-75 (Arg-Leu) in the α-chain. Afteractivation, the 11.2 kD, 74 amino acid peptide C5a from theamino-terminus portion of the α-chain is released. Both C5a and C₃a arepotent stimulators of neutrophils and monocytes (Schindler, R. et al.,Blood 76: 1631-1638 (1990); Haeffner—Cavaillon, N. et al., J. Immunol.138: 794-700 (1987); Cavaillon, J. M. et al., Eur. J. Immunol. 20:253-257 (1990)).

In addition to its anaphylatoxic properties, C5a induces chemotacticmigration of neutrophils (Ward, P. A. et al., J. Immunol. 102: 93-99(1969)), eosinophils (Kay, A. B. et al., Immunol. 24: 969-976 (1973)),basophils (Lett-Brown, M. A. et al., J. Immunol. 117: 246-252 1976)),and monocytes (Snyderman, R. et al., Proc. Soc. Exp. Biol. Med. 138:387-390 1971)). Both C5a and C₅b-9 activate endothelial cells to expressadhesion molecules essential for sequestration of activated leukocytes,which mediate tissue inflammation and injury (Foreman, K. E. et al., J.Clin. Invest. 94: 1147-1155 (1994); Foreman, K. E. et al., Inflammation20: 1-9 (1996); Rollins, S. A. et al., Transplantation 69: 1959-1967(2000)). C5a also mediates inflammatory reactions by causing smoothmuscle contraction, increasing vascular permeability, inducing basophiland mast cell degranulation and inducing release of lysosomal proteasesand oxidative free radicals (Gerard, C. et al., Ann. Rev. Immunol. 12:775-808 (1994)). Furthermore, C5a modulates the hepatic acute-phase geneexpression and augments the overall immune response by increasing theproduction of TNF-α, IL-1-β, IL-6, IL-8, prostaglandins and leukotrienes(Lambris, J. D. et al., In: The Human Complement System in Health andDisease, Volanakis, J. E. ed., Marcel Dekker, New York, pp. 83-118).

The anaphylactic and chemotactic effects of C5a are believed to bemediated through its interaction with the C5a receptor. The human C5areceptor (C5aR) is a 52 kD membrane bound G protein-coupled receptor,and is expressed on neutrophils, monocytes, basophils, eosinophils,hepatocytes, lung smooth muscle and endothelial cells, and renalglomerular tissues (Van-Epps, D. E. et al., J. Immunol. 132: 2862-2867(1984); Haviland, D. L. et al., J. Immunol. 154:1861-1869 (1995);Wetsel, R. A., Immunol. Leff. 44: 183-187 (1995); Buchner, R. R. et al.,J. Immunol. 155: 308-315 (1995); Chenoweth, D. E. et al., Proc. Natl.Acad. Sci. 75: 3943-3947 (1978); Zwirner, J. et al., Mol. Immunol.36:877-884 (1999)). The ligand-binding site of C5aR is complex andconsists of at least two physically separable binding domains. One bindsthe C5a amino terminus (amino acids 1-20) and disulfide-linked core(amino acids 21-61), while the second binds the C5a carboxy-terminal end(amino acids 62-74) (Wetsel, R. A., Curr. Opin. Immunol. 7: 48-53(1995)).

C5a plays important roles in inflammation and tissue injury. Incardiopulmonary bypass and hemodialysis, C5a is formed as a result ofactivation of the alternative complement pathway when human blood makescontact with the artificial surface of the heart-lung machine or kidneydialysis machine (Howard, R. J. et al., Arch. Surg. 123: 1496-1501(1988); Kirklin, J. K. et al., J. Cardiovasc. Surg. 86: 845-857 (1983);Craddock, P. R. et al., N. Engl. J. Med. 296: 769-774 (1977)). C5acauses increased capillary permeability and edema, bronchoconstriction,pulmonary vasoconstriction, leukocyte and platelet activation andinfiltration to tissues, in particular the lung (Czermak, B. J. et al.,J. Leukoc. Biol. 64: 40-48 (1998)). Administration of an anti—C5amonoclonal antibody was shown to reduce cardiopulmonary bypass andcardioplegia-induced coronary endothelial dysfunction (Tofukuji, M. etal., J. Thorac. Cardiovasc. Surg. 116: 1060-1068 (1998)).

C5a is also involved in acute respiratory distress syndrome (ARDS),Chronic Obstructive Pulmonary Disorder (COPD) and multiple organ failure(MOF) (Hack, C. E. et al., Am. J. Med. 1989: 86: 20-26; Hammerschmidt DEet al. Lancet 1980; 1: 947-949; Heideman M. et al. J. Trauma 1984; 4:1038-1043; Marc, MM, et al., Am. J. Respir. Cell and Mol. Biol., 2004:31: 216-219). C5a augments monocyte production of two importantpro-inflammatory cytokines, TNF-α and IL-1. C5a has also been shown toplay an important role in the development of tissue injury, andparticularly pulmonary injury, in animal models of septic shock(Smedegard Get al. Am. J. Pathol. 1989; 135: 489-497; Markus, S., etal., FASEB Journal (2001), 15: 568-570). In sepsis models using rats,pigs and non-human primates, anti—C5a antibodies administered to theanimals before treatment with endotoxin or E. coli resulted in decreasedtissue injury, as well as decreased production of IL-6 (Smedegard, G. etal., Am. J. Pathol. 135: 489-497 (1989); Hopken, U. et al., Eur. J.Immunol. 26: 1103-1109 (1996); Stevens, J. H. et al., J. Clin. Invest.77: 1812-1816 (1986)). More importantly, blockade or C5a with anti—C5apolyclonal antibodies has been shown to significantly improve survivalrates in a caecal ligation/puncture model of sepsis in rats (Czermak,B.J. et al., Nat. Med. 5: 788-792 (1999)). This model share many aspectsof the clinical manifestation of sepsis in humans. (Parker, S.J. et al.,Br. J. Surg. 88: 22-30 (2001)). In the same sepsis model, anti—C5aantibodies were shown to inhibit apoptosis of thymocytes (Guo, R. F. etal., J. Clin. Invest. 106: 1271-1280 (2000)) and prevent MOF(Huber-Lang, M. et al., J. Immunol. 166: 1193-1199 (2001)). Anti—C₅aantibodies were also protective in a cobra venom factor model of lunginjury in rats, and in immune complex-induced lung injury (Mulligan, M.S. et al. J. Clin. Invest. 98: 503-512 (1996)). The importance of C5a inimmune complex-mediated lung injury was later confirmed in mice (Bozic,C. R. et al., Science 26: 1103-1109 (1996)).

C5a is found to be a major mediator in myocardial ischemia-reperfusioninjury. Complement depletion reduced myocardial infarct size in mice(Weisman, H. F. et al., Science 249: 146-151 (1990)), and treatment withanti—C5a antibodies reduced injury in a rat model of hindlimbischemia-reperfusion (Bless, N. M. et al., Am. J. Physiol. 276: L57-L63(1999)). Reperfusion injury during myocardial infarction was alsomarkedly reduced in pigs that were retreated with a monoclonal anti—C5aIgG (Amsterdam, E. A. et al., Am. J. Physiol. 268:H₄₄₈-H₄₅₇ (1995)). Arecombinant human C5aR antagonist reduces infarct size in a porcinemodel of surgical revascularization (Riley, R. D. et al., J. Thorac.Cardiovasc. Surg. 120: 350-358 (2000)).

C5a driven neutrophils also contribute to many bullous diseases (e.g.,bullous pemphigoid, pemphigus vulgaris and pemphigus foliaceus). Theseare chronic and recurring inflammatory disorders clinicallycharacterized by sterile blisters that appear in the sub-epidermal spaceof the skin and mucosa. While autoantibodies to keratinocytes located atthe cutaneous basement membranes are believed to underlie the detachmentof epidermal basal keratinocytes from the underlying basement membrane,blisters are also characterized by accumulation of neutrophils in boththe upper dermal layers and within the blister cavities. In experimentalmodels a reduction of neutrophils or absence of complement (total orC₅-selective) can inhibit formation of sub-epidermal blisters, even inthe presence of high auto-antibody titers.

Complement levels are elevated in patients with rheumatoid arthritis(Jose, P. J. et al., Ann. Rheum. Dis. 49: 747-752 (1990); Grant, E. P.,et al., J. of Exp. Med., 196(11): 1461-1471, (2002)), lupus nephritis(Bao, L., et al., Eur. J. of Immunol., 35(8), 2496-2506, (2005)) andsystemic lupus erythematosus (SLE) (Porcel, J. M. et al., Clin. Immunol.Immunopathol. 74: 283-288 (1995)). C5a levels correlate with theseverity of the disease state. Collagen-induced arthritis in mice andrats resembles the rheumatoid arthritic disease in human. Mice deficientin the C5a receptor demonstrated a complete protection from arthritisinduced by injection of monoclonal anti-collagen Abs (Banda, N. K., etal., J. of Immunol., 2003, 171: 2109-2115). Therefore, inhibition of C5aand/or C5a receptor (C5aR) could be useful in treating these chronicdiseases.

The complement system is believed to be activated in patients withinflammatory bowel disease (IBD) and is thought to play a role in thedisease pathogenesis. Activated complement products were found at theluminal face of surface epithelial cells, as well as in the muscularismucosa and submucosal blood vessels in IBD patients (Woodruff, T. M., etal., J of lmmunol., 2003, 171: 5514-5520).

C5aR expression is upregulated on reactive astrocytes, microglia, andendothelial cells in an inflamed human central nervous system (Gasque,P. et al., Am. J Pathol. 150: 31-41 (1997)). C5a might be involved inneurodegenerative diseases, such as Alzheimer disease (Mukherjee, P. etal., J Neuroimmunol. 105: 124-130 (2000); O'Barr, S. et al., JNeuroimmunol. (2000) 105: 87-94; Farkas, I., et al. J. Immunol. (2003)170:5764-5771), Parkinson's disease, Pick disease and transmissiblespongiform encephalopathies. Activation of neuronal C5aR may induceapoptosis (Farkas I et al. J. Physiol. 1998; 507: 679-687). Therefore,inhibition of C5a and/or C5aR could also be useful in treatingneurodegenerative diseases.

There is some evidence that C5a production worsens inflammationassociated with atopic dermatitis (Neuber, K., et al., Immunology73:83-87, (1991)), and chronic urticaria (Kaplan, A.P., J. Allergy Clin.Immunol. 114; 465-474, (2004).

Psoriasis is now known to be a T cell-mediated disease (Gottlieb, E. L.et al., Nat. Med. 1: 442-447 (1995)). However, neutrophils and mastcells may also be involved in the pathogenesis of the disease (Terui, T.et al., Exp. Dermatol. 9: 1-10; 2000); Werfel, T. et al., Arch.Dermatol. Res. 289: 83-86 (1997)). Neutrophil accumulation under thestratum corneum is observed in the highly inflamed areas of psoriaticplaques, and psoriatic lesion (scale) extracts contain highly elevatedlevels of C5a and exhibit potent chemotactic activity towardsneutrophils, an effect that can be inhibited by addition of a C5aantibody. T cells and neutrophils are chemo-attracted by C5a (Nataf, S.et al., J. Immunol. 162: 4018-4023 (1999); Tsuji, R. F. et al., J.Immunol. 165: 1588-1598 (2000); Cavaillon, J. M. et al., Eur. J Immunol.20: 253-257 (1990)). Additionally expression of C5aR has beendemonstrated in plasmacytoid dendritic cells (pDC) isolated from lesionsof cutaneous lupus erythematous and these cells were shown to displaychemotactic behavior towards C5a, suggesting that blockade of C5aR onpDC might be efficacious in reducing pDC infiltration into inflamed skinin both SLE and psoriasis. Therefore C5a could be an importanttherapeutic target for treatment of psoriasis.

Immunoglobulin G-containing immune complexes (IC) contribute to thepathophysiology in a number of autoimmune diseases, such as systemiclupus erthyematosus, rheumatoid arthritis, Sjogren's disease,Goodpasture's syndrome, and hypersensitivity pneumonitis (Madaio, M. P.,Semin. Nephrol. 19: 48-56 (1999); Korganow, A. S. et al., Immunity 10:451-459 (1999); Bolten, W. K., Kidney Int. 50: 1754-1760 (1996); Ando,M. et al., Curr. Opin. Pulm. Med. 3: 391-399 (1997)). These diseases arehighly heterogeneous and generally affect one or more of the followingorgans: skin, blood vessels, joints, kidneys, heart, lungs, nervoussystem and liver (including cirrhosis and liver fibrosis). The classicalanimal model for the inflammatory response in these IC diseases is theArthus reaction, which features the infiltration of polymorphonuclearcells, hemorrhage, and plasma exudation (Arthus, M., C. R. Soc. Biol.55: 817-824 (1903)). Recent studies show that C5aR deficient mice areprotected from tissue injury induced by IC (Kohl, J. et al., Mol.Immunol. 36: 893-903 (1999); Baumann, U. et al., J. Immunol. 164:1065-1070 (2000)). The results are consistent with the observation thata small peptidic anti—C5aR antagonist inhibits the inflammatory responsecaused by IC deposition (Strachan, A. J. et al., J. Immunol. 164:6560-6565 (2000)). Together with its receptor, C5a plays an importantrole in the pathogenesis of IC diseases. Inhibitors of C5a and C5aRcould be useful to treat these diseases.

Descripton of Related Art

Non-peptide based C5a receptor antagonist have been reported as beingeffective for treating endotoxic shock in rats (Stracham, A. J., et al.,J. of Immunol. (2000), 164(12): 6560-6565); and for treating IBD in arat model (Woodruff, T. M., et al., J of Immunol., 2003, 171:5514-5520). Non-peptide based C5a receptor modulators also have beendescribed in the patent literature by Neurogen Corporation, (e.g.,WO2004/043925, WO2004/018460, WO2005/007087, WO03/082826, WO03/08828,WO02/49993, WO03/084524); Dompe S.P.A. (WO02/029187); The University ofQueenland (WO2004/100975); and ChemoCentryx (WO2010/075257).

There is considerable experimental evidence in the literature thatimplicates increased levels of C5a with a number of diseases anddisorders, in particular in autoimmune and inflammatory diseases anddisorders. Thus, there remains a need in the art for new small organicmolecule modulators, e.g., agonists, preferably antagonists, partialagonists, of the C5a receptor (C5aR) that are useful for inhibitingpathogenic events, e.g., chemotaxis, associated with increased levelsanaphylatoxin activity. The present invention fulfills this and otherneeds.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention provide compounds of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein the symbols,letters and subscripts n, m, a, b, e, X¹, R¹, R^(2a), R^(2b), R³, R⁴ andR⁵ have the meanings provided in the description below.

In addition to the compounds provided herein, the present inventionfurther provides pharmaceutical compositions containing one or more ofthese compounds, as well as methods for the use of these compounds intherapeutic methods, primarily to treat diseases associated C5asignaling activity.

In yet another aspect, the present invention provides methods ofdiagnosing disease in an individual. In these methods, the compoundsprovided herein are administered in labeled form to a subject, followedby diagnostic imaging to determine the presence or absence of C5aRand/or the localization of cells expressing a C5aR receptor. In arelated aspect, a method of diagnosing disease is carried out bycontacting a tissue or blood sample with a labeled compound as providedherein and determining the presence, absence, amount, or localization ofC5aR in the sample.

BRIEF DESCRIPTION OF THE DRAWINGS

NOT APPLICABLE

DETAILED DESCRIPTION OF THE INVENTION I. ABBREVIATION AND DEFINITIONS

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), isobutenyl,2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1— and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkyl” refers to a cycloalkyl group that contain from one tofive heteroatoms selected from N, O, and S, wherein the nitrogen andsulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. The heterocycloalkyl may be a monocyclic, abicyclic or a polycylic ring system. Non limiting examples ofheterocycloalkyl groups include pyrrolidine, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrhydrothiophene, quinuclidine, and the like. Aheterocycloalkyl group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

The term “heteroalkyl,” by itself or in combination with another term,means, unless otherwise stated, a stable straight or branched chain, orcyclic hydrocarbon radical, or combinations thereof, consisting of thestated number of carbon atoms and from one to three heteroatoms selectedfrom the group consisting of O, N, Si and S, and wherein the nitrogenand sulfur atoms may optionally be oxidized and the nitrogen heteroatommay optionally be quaternized. The heteroatom(s) O, N and S may beplaced at any interior position of the heteroalkyl group. The heteroatomSi may be placed at any position of the heteroalkyl group, including theposition at which the alkyl group is attached to the remainder of themolecule. Examples include —CH₂—CH₂—O—CH₃, —CH₂—CH₂—NH—CH₃,—CH₂—CH₂—N(CH₃)—CH₃, —CH₂—S—CH₂—CH₃, —CH₂—CH₂,—S(O)—CH₃,—CH₂—CH₂—S(O)₂—CH₃, —CH═CH—O—CH₃, —Si(CH₃)₃, —CH₂—CH═N—OCH₃, and—CH═CH—N(CH₃)—CH₃. Up to two heteroatoms may be consecutive, such as,for example, —CH₂—NH—OCH₃ and —CH₂—O—Si(CH₃)₃. Similarly, the terms“heteroalkenyl” and “heteroalkynyl” by itself or in combination withanother term, means, unless otherwise stated, an alkenyl group oralkynyl group, respectively, that contains the stated number of carbonsand having from one to three heteroatoms selected from the groupconsisting of O, N, Si and S, and wherein the nitrogen and sulfur atomsmay optionally be oxidized and the nitrogen heteroatom may optionally bequaternized. The heteroatom(s) O, N and S may be placed at any interiorposition of the heteroalkyl group.

The term “heteroalkylene” by itself or as part of another substituentmeans a divalent radical, saturated or unsaturated or polyunsaturated,derived from heteroalkyl, as exemplified by —CH₂—CH₂—S—CH₂CH₂— and—CH₂—S—CH₂—CH₂—NH—CH₂—, —O—CH₂—CH═CH—, —CH₂—CH═C(H)CH₂—O—CH₂— and—S—CH₂—C—. For heteroalkylene groups, heteroatoms can also occupy eitheror both of the chain termini (e.g., alkyleneoxy, alkylenedioxy,alkyleneamino, alkylenediamino, and the like).

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as —NR^(a)R^(b) ismeant to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl andthe like.

The term “hydroxyalkyl” is used in its conventional sense, and refers tobranched or straight chain alkyl group substituted with at least onehydroxyl group. The hydroxyl group may be at any position in the alkylgroup. For example, the term “C₁₋₄hydroxylalkyl” is meant to includehydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxyisopropyl, and thelike.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalaziniyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzooxazolyl, benzotriazolyl,benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl,thienopyridinyl, thienopyrimidinyl, pyrazolopyrimidinyl, pyrrolopyridyl,imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl,quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl,imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl,pyrrolyl, thiazolyl, furyl, thienyl and the like. Substituents for eachof the above noted aryl and heteroaryl ring systems are selected fromthe group of acceptable substituents described below.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occuring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. The compounds of the present invention may alsocontain unnatural proportions of atomic isotopes at one or more of theatoms that constitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule.

II. DESCRIPTION OF THE EMBODIMENTS

A. Compounds

In one aspect, the present invention provides compounds of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein,

-   -   ring vertex a is N or C(R^(2c)), ring vertex b is N or        C(R^(2d)), and ring vertex e is N or C(R^(2e)), wherein no more        than one of a, b and e is N;    -   X¹ is selected from the group consisting of a bond,        C₁₋₁₈alkylene, C(O), C(O)—C₁₋₄alkylene, and S(O)₂;    -   R¹ is selected from the group consisting of        -   a) 5- to 10-membered heteroaryl having from 1 to 4            heteroatoms as ring vertices selected from N, O and S;        -   b) C₆₋₁₀ aryl;        -   c) C₃₋₈ cycloalkyl;        -   d) 4- to 8-membered heterocycloalkyl having from 1 to 2            heteroatoms as ring vertices selected from N, O and S; and        -   e) C₁₋₈alkyl, C₁₋₈alkoxy, C₁₋₈haloalkyl, —C(O)NR^(1a)R^(1b),            and —CO₂R^(1a); wherein R^(1a) and            -   R^(1b) are each independently selected from the group                consisting of hydrogen, C₁₋₈alkyl, C₆₋₁₀ aryl, and                —C₁₋₆alkylene-C₆₋₄₀aryl;

wherein the group —X¹—R¹ is optionally substituted with 1 to 5 R^(x)substituents;

-   -   R^(2a) and R^(2e) are each independently selected from the group        consisting of hydrogen, C₁₋₆ alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl,        —O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl, —C₁₋₆alkyl-O—C₁₋₆ alkyl,        —C₁₋₆alkyl-S—C₁₋₆alkyl, CN, and halogen, and at least one of        R^(2a) and R^(2e) is other than hydrogen;    -   R^(2b), R^(2c), and R^(2d) are each independently selected from        the group consisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy,        C₁₋₆haloalkyl, —O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl,        —C₁₋₆alkyl-O—C₁₋₆alkyl, —C₁₋₆alkyl-S—C₁₋₆alkyl, cyano, and        halogen;    -   each R³ is independently selected from the group consisting of        hydroxyl, C₁₋₄alkyl, C₁₋₄haloalkyl and C₁₋₄hydroxyalkyl, and        optionally two R³ groups on the same carbon atom are combined to        form oxo (═O), and optionally two R³ groups and the carbon atoms        they are attached to form a 3-6 membered ring with 0-2        hetereoatoms as ring members selected from O, N, and S;    -   R⁴ is independently selected from the group consisting of        —X²—OR^(4a), —X²—NR^(4a)R^(4b), —X²—CONR^(4a)R^(4b),        —X²—NR^(4a)—C(O)R^(4a), —X²—NR^(4a)—C(O)NR^(4a)R^(4b),        —X²NR^(4a)—C(O)OR^(4a), —X²—NR^(4a)—C(O)—C₁₋₃alkylene-OR^(4a)        and —X²—NR^(4a)—C(O)—C₁₋₃alkylene-NR^(4a)R^(4b); wherein each X²        is independently a bond, C(O), C₁₋₄alkylene, C(O)—C₁₋₄alkylene,        and C₁₋₄alkylene-C(O), and each R^(4a) and R^(4b) is        independently selected from the group consisting of hydrogen,        C₁₋₄alkyl, and C₁₋₄haloalkyl;    -   each R⁵ is independently selected from the group consisting of        C₁₋₈alkyl, C₁₋₈alkoxy, C₁₋₈haloalkyl, C₁₋₈haloalkoxy,        C₁₋₈hydroxyalkyl, halogen, OH, CN, C(O)R^(5a) and CO₂R^(5a);        wherein each R^(5a) is independently selected from the group        consisting of hydrogen, C₁₋₄alkyl, and C₁₋₄haloalkyl;    -   each R^(x) is independently selected from the group consisting        of halogen, CN, C₁₋₄alkyl, C₁₋₄alkoxy, C₁₋₄haloalkyl,        C₁₋₄haloalkoxy, C₁₋₄hydroxy, C₂-4alkenyl, C₃₋₆cycloalkyl,        CO₂-C₁₋₄alkyl, and CONH₂;    -   the subscript m is 0, 1, 2, 3 or 4; and    -   the subscript n is 0, 1, 2 or 3.

In one group of embodiments for the compounds of formula (I), R⁴ isselected from the group consisting of

In another group of embodiments for the compounds of formula (I), or apharmaceutically acceptable salt thereof, R⁴ is selected from the groupconsisting of

In yet another group of embodiments for the compounds of formula (I), ora pharmaceutically acceptable salt thereof, wherein R⁴ is selected fromthe group consisting of

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups of embodimentsnoted above, in certain selected embodiments, X¹ is a bond; in otherselected embodiments, X¹ is C(O); in still other selected embodiments,X¹ is C₁₋₈alkylene; in yet other selected embodiments, X¹ isC(O)—C₁₋₄alkylene or S(O)₂.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, wherein R¹ is a 5-to 10-membered heteroaryl having from 1 to 4 heteroatoms as ringvertices selected from N, O and S; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents. In still furtherembodiments, R¹ is selected from the group consisting of pyrazolyl,pyridyl, pyrimidinyl, imidazolyl, thiazolyl, thiadiazolyl and pyrazinyl;and wherein the group —X¹—R¹ is optionally substituted with 1 to 4 R^(x)substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, wherein R¹ isC₆₋₁₀aryl; and wherein the group is optionally substituted with 1 to 4R^(x) substituents. In still further embodiments, R¹ is phenyl; andwherein the group —X¹-R¹ is optionally substituted with 1 to 4 R^(x)substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, R¹ is C₃₋₈aryl;and wherein the group —X¹—R¹ is optionally substituted with 1 to 4 R^(x)substituents. In still further embodiments, R¹ is selected from thegroup consisting of cyclobutyl, cyclopentyl and cyclohexyl; and whereinthe group —X¹—R¹ is optionally substituted with 1 to 4 R^(x)substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, R¹ is a 4- to8-membered heterocycloalkyl having from 1 to 2 heteroatoms as ringvertices selected from N, O and S; and wherein the group is optionallysubstituted with 1 to 4 R^(x) substituents. In still further selectedembodiments, R¹ is selected from the group consisting of oxetanyl,tetrahydrofuranyl, tetrahydropyranyl and morpholinyl; and wherein thegroup is optionally substituted with 1 to 4 R^(x) substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, R¹ is selectedfrom the group consisting of C₁₋₈alkyl, C₁₋₈alkoxy, C₁₋₈haloalkyl,—C(O)NR^(1a)R^(1b), and —CO₂R^(1a); wherein R^(1a) and R^(1b) are eachindependently selected from the group consisting of hydrogen, C₁₋₈alkyl,C₆₋₁₀aryl, and —C₁₋₆alkylene-C₆₋₁₀aryl; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the groups or selectedembodiments noted above, in some further embodiments, R¹ is selectedfrom the group consisting of phenyl, pyridyl, pyrimidinyl, andpyrazinyl; and wherein the group —X¹—R¹ is optionally substituted with 1to 4 R^(x) substituents.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, ring vertices a and b are CH; R^(2b) is H;ring vertex e is C(R^(2e)), and R^(2a) and R^(2e) are independentlyselected from the group consisting of C₁₋₆alkyl, C₁₋₆alkoxy,C₁₋₆haloalkyl,—O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl, —C₁₋₆alkyl-O—C₁₋₆alkyl,—C₁₋₆alkyl-S—C₁₋₆alkyl, CN, and halogen.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, ring vertices a and b are CH; R^(2b) is H;ring vertex e is C(R^(2e)), and R^(2a) and R^(2e) are independentlyselected from the group consisting of C₁₋₆alkyl, C₁₋₆alkoxy and halogen.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, the subscript n is 0, 1 or 2 and each R⁵,when present, is selected from the group consisting of F, Cl, CN,C₁₋₄alkyl and C₁₋₄alkoxy. In still further selected embodiments, thesubscript n is 0, 1 or 2 and each R⁵, when present, is selected from thegroup consisting of F, Cl, CN, CH₃ and OCH₃.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, the subscript m is 0, 1 or 2 and each R³,when present, is C₁₋₄alkyl.

In a particular group of embodiments of the compounds of formula (I), ora pharmaceutically acceptable salt thereof, R¹ is selected from thegroup consisting of phenyl or pyridyl, wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents; ring vertices aand b are CH; R^(2b) is H; ring vertex e is C(R^(2e)), and R^(2a) andR^(2e) are independently selected from the group consisting ofC₁₋₆alkyl, C₁₋₆alkoxy and halogen; m is 0, 1 or 2 and each R³, whenpresent, is CH₃, R⁴ is selected from the group consisting of

n is 0, 1 or 2 and each R⁵, when present, is selected from the groupconsisting of F, Cl, CN, CH₃ and OCH₃.

In some embodiments of the compounds of formula (I), or apharmaceutically acceptable salt thereof, R¹ is selected from the groupconsisting of

In some embodiments of the compounds of formula (I), or apharmaceutically acceptable salt thereof, —X—R¹ is selected from thegroup consisting of:

In some embodiments of the compounds of formula (I), or apharmaceutically acceptable salt thereof, R¹ is selected from the groupconsisting of:

In some embodiments of the compounds of formula (I), or apharmaceutically acceptable salt thereof, R¹ is selected from the groupconsisting of:

In some embodiments of the compounds of formula (I), or apharmaceutically acceptable salt thereof, R¹ is selected from the groupconsisting of:

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, the group

is selected from the group consisting of

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, wherein n is 0.

With reference to the compounds of formula (I), or a pharmaceuticallyacceptable salt thereof, as well as any of the embodiments noted above,in some further embodiments, the subscript n is 2 and the two R³ groupsare on the same carbon atom and are combined to form oxo (═O).

In some selected embodiments, provided herein is a compound selectedfrom the group consisting of:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of Formula (I) is a compound describedin the Examples section and the accompanying Tables.

Preparation of Compounds

Certain compounds of the invention can be prepared following methodologyas described in the Examples section of this document. In addition, thesyntheses of certain intermediate compounds that are useful in thepreparation of compounds of the invention are also described.

B. Pharmaceutical Compositions

In addition to the compounds provided above, compositions for modulatingC5a activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self emulsifications as described in U.S. PatentApplication 2002-0012680, hard or soft capsules, syrups, elixirs,solutions, buccal patch, oral gel, chewing gum, chewable tablets,effervescent powder and effervescent tablets. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents, antioxidants andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as cellulose, silicon dioxide, aluminumoxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example PVP, cellulose, PEG, starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated,enterically or otherwise, by known techniques to delay disintegrationand absorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate may beemployed. They may also be coated by the techniques described in theU.S. Pat. Nos. 4,256,108; 4,166,452; and 4,265,874 to form osmotictherapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols. Additionally, the compounds can be administeredvia ocular delivery by means of solutions or ointments. Still further,transdermal delivery of the subject compounds can be accomplished bymeans of iontophoretic patches and the like. For topical use, creams,ointments, jellies, solutions or suspensions, etc., containing thecompounds of the present invention are employed. As used herein, topicalapplication is also meant to include the use of mouth washes andgargles.

The compounds of this invention may also be coupled a carrier that is asuitable polymers as targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer,polyhydroxy-propyl-methacrylamide-phenol,polyhydroxyethyl-aspartamide-phenol, or polyethyleneoxide-polylysinesubstituted with palmitoyl residues. Furthermore, the compounds of theinvention may be coupled to a carrier that is a class of biodegradablepolymers useful in achieving controlled release of a drug, for examplepolylactic acid, polyglycolic acid, copolymers of polylactic andpolyglycolic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates andcross linked or amphipathic block copolymers of hydrogels. Polymers andsemipermeable polymer matrices may be formed into shaped articles, suchas valves, stents, tubing, prostheses and the like. In one embodiment ofthe invention, the compound of the invention is coupled to a polymer orsemipermeable polymer matrix that is formed as a stent or stert-graftdevice.

The pharmaceutical compositions of the present disclosure may beformulated with one or more additional therapeutic agents. The one ormore additional therapeutic agent is selected from the group consistingof corticosteroids, steroids, immunosuppressants, Immunoglobulin Gagonists, Dipeptidyl peptidase IV inhibitors, Lymphocyte functionantigen-3 receptor antagonists, Interleukin-2 ligands, Interleukin-1beta ligand inhibitors, IL-2 receptor alpha subunit inhibitors, HGF genestimulators, IL-6 antagonists, IL-5 antagonists, Alpha 1 antitrypsinstimulators, Cannabinoid receptor antagonists, Histone deacetylaseinhibitors, AKT protein kinase inhibitors, CD20 inhibitors, Abl tyrosinekinase inhibitors, JAK tyrosine kinase inhibitors, TNF alpha ligandinhibitors, Hemoglobin modulators, TNF antagonists, proteasomeinhibitors, CD3 modulators, Hsp 70 family inhibitors, Immunoglobulinagonists, CD30 antagonists, tubulin antagonists, Sphingosine-1-phosphatereceptor-1 agonists, connective tissue growth factor ligand inhibitors,caspase inhibitors, adrenocorticotrophic hormone ligands, Btk tyrosinekinase inhibitors, Complement Cls subcomponent inhibitors,Erythropoietin receptor agonists, B-lymphocyte stimulator ligandinhibitors, Cyclin-dependent kinase-2 inhibitors, P-selectinglycoprotein ligand-1 stimulators, mTOR inhibitors, Elongation factor 2inhibitors, Cell adhesion molecule inhibitors, Factor XIII agonists,Calcineurin inhibitors, Immunoglobulin G1 agonists, Inosinemonophosphate dehydrogenase inhibitors, Complement Cls subcomponentinhibitors, Thymidine kinase modulators, Cytotoxic T-lymphocyteprotein-4 modulators, Angiotensin II receptor antagonists, AngiotensinII receptor modulators, TNF superfamily receptor 12A antagonists, CD52antagonists, Adenosine deaminase inhibitors, T-cell differentiationantigen CD6 inhibitors, FGF-7 ligands, dihydroorotate dehydrogenaseinhibitors, Syk tyrosine kinase inhibitors, Interferon type I receptorantagonists, Interferon alpha ligand inhibitors, Macrophage migrationinhibitory factor inhibitors, Integrin alpha-V/beta-6 antagonists,Cysteine protease stimulators, p38 MAP kinase inhibitors, TP53 geneinhibitors, Shiga like toxin I inhibitors, Fucosyltransferase 6stimulators, Interleukin 22 ligands, IRS1 gene inhibitors, Proteinkinase C stimulators, Protein kinase C alpha inhibitors, CD74antagonists, Immunoglobulin gamma Fc receptor IIB antagonists, T-cellantigen CD7 inhibitors, CD95 antagonists, N acetylmannosamine kinasestimulators, Cardiotrophin-1 ligands, Leukocyte elastase inhibitors,CD40 ligand receptor antagonists, CD40 ligand modulators, IL-17antagonists, TLR-2 antagonists, Mannan-binding lectin serine protease-2(MASP-2) inhibitors, Factor B inhibitors, Factor D inhibitors, C₃aRmodulators, C5aR² modulators, T cell receptor antagonists, PD-1inhibitors, PD-L1 inhibitors, TIGIT inhibitors, TIM-3 inhibitors, LAG-3inhibitors, VISTA inhibitors, STING agonists, IDO inhibitors, adenosinereceptor modulators, CD39 inhibitors, CD73 inhibitors, antagonists ofthe chemokine receptors, especially CXCR1, CXCR2, CXCR3, CXCR4, CXCR7,CCR1, CCR2, CCR³, CCR4, CCR5, CCR7, CCR7, CCR9, CX3CR1 and CXCR6, andcombinations thereof.

In some embodiments, the one or more additional therapeutic agent isselected from the group consisting of obinutuzumab, rituximab,ocrelizumab, cyclophosphamide, prednisone, hydrocortisone,hydrocortisone acetate, cortisone acetate, tixocortol pivalate,prednisolone, methylprednisolone, triamcinolone acetonide, triamcinolonealcohol, mometasone, amcinonide, budesonide, desonide, fluocinonide,fluocinolone acetonide, halcinonide, betamethasone, betamethasone sodiumphosphate, dexamethasone, dexamethasone sodium phosphate, fluocortolone,hydrocortisone-17-valerate, halometasone, alclometasone dipropionate,beclomethasone, betamethasone valerate, betamethasone dipropionate,prednicarbate, clobetasone-17-butyrate, clobetasol-17-propionate,fluocortolone caproate, fluocortolone pivalate, fluprednidene acetate,hydrocortisone-17-butyrate, hydrocortisone-17-aceponate,hydrocortisone-17-buteprate, ciclesonide and prednicarbate, GB-0998,immuglo, begelomab, alefacept, aldesleukin, gevokizumab, daclizumab,basiliximab, inolimomab, beperminogene perplasmid, sirukumab,tocilizumab, clazakizumab, mepolizumab, fingolimod, panobinostat,triciribine, nilotinib, imatinib, tofacitinib, momelotinib, peficitinib,itacitinib, infliximab, PEG-bHb-CO, etanercept, ixazomib, bortezomib,muromonab, otelixizumab, gusperimus, brentuximab vedotin, Ponesimod,KRP-203, FG-3019, emricasan, corticotropin, ibrutinib, cinryze,conestat, methoxy polyethylene glycol-epoetin beta, belimumab,blisibimod, atacicept, seliciclib, neihulizumab, everolimus, sirolimus,denileukin diftitox, LMB-2, natalizumab, catridecacog, ciclosporin,tacrolimus, voclosporin, voclosporin, canakinumab, mycophenolate,mizoribine, CE-1145, TK-DLI, abatacept, belatacept, olmesartanmedoxomil, sparsentan, TXA-127, BIIB-023, alemtuzumab, pentostatin,itolizumab, palifermin, leflunomide, PRO-140, cenicriviroc,fostamatinib, anifrolumab, sifalimumab, BAX-069, BG-00011, losmapimod,QPI-1002, ShigamAbs, TZ-101, F-652, reparixin, ladarixin, PTX-9908,aganirsen, APH-703, sotrastaurin, sotrastaurin, milatuzumab, SM-101,T-Guard, APG-101, DEX-M74, cardiotrophin-1, tiprelestat, ASKP-1240,BMS-986004, HPH-116, KD-025, OPN-305, TOL-101, defibrotide,pomalidomide, Thymoglobulin, laquinimod, remestemcel-L, Equineantithymocyte immunoglobulin, Stempeucel, LIV-Gamma, Octagam 10%,t2c-001, 99mTc-sestamibi, Clairyg, Prosorba, pomalidomide, laquinimod,teplizumab, FCRx, solnatide, foralumab, ATIR-101, BPX-501, ACP-01,ALLO-ASC-DFU, irbesartan+propagermanium, ApoCell, cannabidiol, RGI-2001,saratin, anti—CD3 bivalent antibody-diphtheria toxin conjugate, NOX-100,LT-1951, OMS721, ALN—CCS, ACH-4471, AMY-101, Acthar gel, andCD4+CD25+regulatory T-cells, MEDI7814, P32, P59, pembrolizumab,nivolumab, atezolizumab, avelumab, durvalumab, CCX354, CCX721, CCX9588,CCX140, CCX872, CCX598, CCX6239, CCX587, CCX624, CCX282, CCX025, CCX507,CCX430, CCX765, CCX758, CCX771, CCX662, CCX650, and combinationsthereof. Further discussions of combination therapy are included in the“Methods of Use” section of this application.

A. Methods of Use

The compounds of the invention may be used as agonists, (preferably)antagonists, partial agonists, inverse agonists, of C5a receptors in avariety of contexts, both in vitro and in vivo. In one embodiment, thecompounds of the invention are C5aR antagonist that can be used toinhibit the binding of C5a receptor ligand (e.g., C5a) to C5a receptorin vitro or in vivo. In general, such methods comprise the step ofcontacting a C5a receptor with a sufficient amount of one or more C5areceptor modulators as provided herein, in the presence of C5a receptorligand in aqueous solution and under conditions otherwise suitable forbinding of the ligand to C5a receptor. The C5a receptor may be presentin suspension (e.g., in an isolated membrane or cell preparation), in acultured or isolated cell, or in a tissue or organ.

Preferably, the amount of C5a receptor modulator contacted with thereceptor should be sufficient to inhibit C5a binding to C5a receptor invitro as measured, for example, using a radioligand binding assay,calcium mobilization assay, or chemotaxis assay as described herein.

In one embodiment of the invention, the C5a modulators of the inventionare used to modulate, preferably inhibit, the signal-transducingactivity of a C5a receptor, for example, by contacting one or morecompound(s) of the invention with a C5a receptor (either in vitro or invivo) under conditions suitable for binding of the modulator(s) to thereceptor. The receptor may be present in solution or suspension, in acultured or isolated cell preparation or within a patient. Anymodulation of the signal transducing activity may be assessed bydetecting an effect on calcium ion calcium mobilization or by detectingan effect on C5a receptor-mediated cellular chemotaxis. In general, aneffective amount of C5a modulator(s) is an amount sufficient to modulateC5a receptor signal transducing activity in vitro within a calciummobilization assay or C5a receptor-mediated cellular chemotaxis within amigration assay.

When compounds of the invention are used to inhibit C5areceptor-mediated cellular chemotaxis, preferably leukocyte (e.g.,neutrophil) chemotaxis, in an in vitro chemotaxis assay, such methodscomprise contacting white blood cells (particularly primate white bloodcells, especially human white blood cells) with one or more compounds ofthe invention. Preferably the concentration is sufficient to inhibitchemotaxis of white blood cells in an in vitro chemotaxis assay, so thatthe levels of chemotaxis observed in a control assay are significantlyhigher, as described above, than the levels observed in an assay towhich a compound of the invention has been added.

In another embodiment, the compounds of the present invention furthercan be used for treating patients suffering from conditions that areresponsive to C5a receptor modulation. As used herein, the term“treating” or “treatment” encompasses both disease-modifying treatmentand symptomatic treatment, either of which may be prophylactic (i.e.,before the onset of symptoms, in order to prevent, delay or reduce theseverity of symptoms) or therapeutic (i.e., after the onset of symptoms,in order to reduce the severity and/or duration of symptoms). As usedherein, a condition is considered “responsive to C5a receptormodulation” if modulation of

C5a receptor activity results in the reduction of inappropriate activityof a C5a receptor. As used herein, the term “patients” include primates(especially humans), domesticated companion animals (such as dogs, cats,horses, and the like) and livestock (such as cattle, pigs, sheep, andthe like), with dosages as described herein.

Conditions that can be Treated by C5a Modulation:

Autoimmune disorders—e.g., Rheumatoid arthritis, systemic lupuserythematosus, Guillain-Barre syndrome, pancreatitis, lupus nephritis,lupus glomerulonephritis, psoriasis, Crohn's disease, vasculitis,irritable bowel syndrome, dermatomyositis, multiple sclerosis, bronchialasthma, dense deposit disease, pemphigus, pemphigoid, scleroderma,myasthenia gravis, autoimmune hemolytic and thrombocytopenic states,Goodpasture's syndrome (and associated glomerulonephritis and pulmonaryhemorrhage), C₃-glomerulopathy, C₃-glomerulonephritis,membranoproliferative glomerulonephritis, Kawasaki disease, IGsnephropathy, immunovasculitis, tissue graft rejection, graft versus hostdisease, hyperacute rejection of transplanted organs; and the like.

Inflammatory disorders and related conditions—e.g., Neutropenia, sepsis,septic shock, Alzheimer's disease, multiple sclerosis, neutrophilia,stroke, inflammatory bowel disease (IBD), inflammation associated withsevere burns, lung injury, and ischemia-reperfusion injury,osteoarthritis, as well as acute (adult) respiratory distress syndrome(ARDS), chronic pulmonary obstructive disorder (COPD), systemicinflammatory response syndrome (SIRS), atopic dermatitis, psoriasis,chronic urticaria and multiple organ dysfunction syndrome (MODS)Hemolytic uremic syndrome, atypical hemolytic uremic syndrome (aHUS).Also included are pathologic sequellae associated with insulin-dependentdiabetes mellitus (including diabetic retinopathy), lupus nephropathy,Heyman nephritis, membranous nephritis and other forms ofglomerulonephritis, contact sensitivity responses, and inflammationresulting from contact of blood with artificial surfaces that can causecomplement activation, as occurs, for example, during extracorporealcirculation of blood (e.g., during hemodialysis or via a heart-lungmachine, for example, in association with vascular surgery such ascoronary artery bypass grafting or heart valve replacement), or inassociation with contact with other artificial vessel or containersurfaces (e.g., ventricular assist devices, artificial heart machines,transfusion tubing, blood storage bags, plasmapheresis,plateletpheresis, and the like). Also included are diseases related toischemia/reperfusion injury, such as those resulting from transplants,including solid organ transplant, and syndromes such as ischemicreperfusion injury, ischemic colitis and cardiac ischemia. Compounds ofthe instant invention may also be useful in the treatment of age-relatedmacular degeneration (Hageman et al, P.N.A.S.102: 7227-7232, 2005).

Cardiovascular and Cerebrovascular Disorders—e.g., myocardialinfarction, coronary thrombosis, vascular occlusion, post-surgicalvascular reocclusion, atherosclerosis, traumatic central nervous systeminjury, and ischemic heart disease. In one embodiment, an effectiveamount of a compound of the invention may be administered to a patientat risk for myocardial infarction or thrombosis (i.e., a patient who hasone or more recognized risk factor for myocardial infarction orthrombosis, such as, but not limited to, obesity, smoking, high bloodpressure, hypercholesterolemia, previous or genetic history ofmyocardial infarction or thrombosis) in order reduce the risk ofmyocardial infarction or thrombosis.

Oncologic Diseases or Disorders—e.g., melanoma, lung cancer, lymphoma,sarcoma, carcinoma, fibrosarcoma, liposarcoma, chondrosarcoma,osteogenic sarcoma, angiosarcoma, lymphangiosarcoma, synovioma,mesothelioma, meningioma, leukemia, lymphoma, leiomyosarcoma,rhabdomyosarcoma, squamous cell carcinoma, basal cell carcinoma,adenocarcinoma, papillary carcinoma, cystadenocarcinoma, bronchogeniccarcinoma, renal cell carcinoma, hepatocellular carcinoma, transitionalcell carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, wilm'stumor, pleomorphic adenoma, liver cell papilloma, renal tubular adenoma,cystadenoma, papilloma, adenoma, leiomyoma, rhabdomyoma, hemangioma,lymphangioma, osteoma, chondroma, lipoma and fibroma.

Diseases of Vasculitis—Vasculitic dseases are characterized byinflammation of the vessels. Infiltration of leukocytes leads todestruction of the vessel walls, and the complement pathway is believedto play a major role in initiating leukocyte migration as well as theresultant damage manifested at the site of inflammation (Vasculitis,Second Edition, Edited by Ball and Bridges, Oxford University Press, pp47-53, 2008). The compounds provided in the present invention can beused to treat leukoclastic vasculitis, Anti-neutrophil cytoplasmicantibody (ANCA) associated vasculitis, immune vasculitis Wegener'sgranulomatosis, microscopic polyangiitis, Churg-Strauss syndrome,Henoch-Schonlein purpura, polyateritis nodosa, Rapidly

Progressive Glomerulonephritis (RPGN), cryoglobulinaemia, giant cellarteritis (GCA), Behcet's disease and Takayasu's arteritis (TAK).

HIV infection and AIDS—C5a receptor modulators provided herein may beused to inhibit HIV infection, delay AIDS progression or decrease theseverity of symptoms or HIV infection and AIDS.

Neurodegenerative disorders and related diseases—Within further aspects,C5a antagonists provided herein may be used to treat Alzheimer'sdisease, multiple sclerosis, and cognitive function decline associatedwith cardiopulmonary bypass surgery and related procedures.

In one embodiment of the invention, the compounds of the invention canbe used for the treatment of diseases selected from the group consistingof sepsis (and associated disorders), COPD, rheumatoid arthritis, lupusnephritis and multiple sclerosis.

Treatment methods provided herein include, in general, administration toa patient an effective amount of one or more compounds provided herein.Suitable patients include those patients suffering from or susceptibleto (i.e., prophylactic treatment) a disorder or disease identifiedherein. Typical patients for treatment as described herein includemammals, particularly primates, especially humans. Other suitablepatients include domesticated companion animals such as a dog, cat,horse, and the like, or a livestock animal such as cattle, pig, sheepand the like.

In general, treatment methods provided herein comprise administering toa patient an effective amount of a compound one or more compoundsprovided herein. In a preferred embodiment, the compound(s) of theinvention are preferably administered to a patient (e.g., a human)orally or topically. The effective amount may be an amount sufficient tomodulate C5a receptor activity and/or an amount sufficient to reduce oralleviate the symptoms presented by the patient. Preferably, the amountadministered is sufficient to yield a plasma concentration of thecompound (or its active metabolite, if the compound is a pro-drug) highenough to detectably inhibit white blood cell (e.g., neutrophil)chemotaxis in vitro. Treatment regimens may vary depending on thecompound used and the particular condition to be treated; for treatmentof most disorders, a frequency of administration of 4 times daily orless is preferred. In general, a dosage regimen of 2 times daily is morepreferred, with once a day dosing particularly preferred. It will beunderstood, however, that the specific dose level and treatment regimenfor any particular patient will depend upon a variety of factorsincluding the activity of the specific compound employed, the age, bodyweight, general health, sex, diet, time of administration, route ofadministration, rate of excretion, drug combination (i.e., other drugsbeing administered to the patient) and the severity of the particulardisease undergoing therapy, as well as the judgment of the prescribingmedical practitioner. In general, the use of the minimum dose sufficientto provide effective therapy is preferred. Patients may generally bemonitored for therapeutic effectiveness using medical or veterinarycriteria suitable for the condition being treated or prevented.

Dosage levels of the order of from about 0.1 mg to about 140 mg perkilogram of body weight per day are useful in the treatment orpreventions of conditions involving pathogenic C5a activity (about 0.5mg to about 7 g per human patient per day). The amount of activeingredient that may be combined with the carrier materials to produce asingle dosage form will vary depending upon the host treated and theparticular mode of administration. Dosage unit forms will generallycontain between from about 1 mg to about 500 mg of an active ingredient.For compounds administered orally, transdermally, intravaneously, orsubcutaneously, it is preferred that sufficient amount of the compoundbe administered to achieve a serum concentration of 5 ng(nanograms)/mL-10 μg (micrograms)/mL serum, more preferably sufficientcompound to achieve a serum concentration of 20 ng-1 μg/ml serum shouldbe administered, most preferably sufficient compound to achieve a serumconcentration of 50 ng/ml-200 ng/ml serum should be administered. Fordirect injection into the synovium (for the treatment of arthritis)sufficient compounds should be administered to achieve a localconcentration of approximately 1 micromolar.

Frequency of dosage may also vary depending on the compound used and theparticular disease treated. However, for treatment of most disorders, adosage regimen of 4 times daily, three times daily, or less ispreferred, with a dosage regimen of once daily or 2 times daily beingparticularly preferred. It will be understood, however, that thespecific dose level for any particular patient will depend upon avariety of factors including the activity of the specific compoundemployed, the age, body weight, general health, sex, diet, time ofadministration, route of administration, and rate of excretion, drugcombination (i.e., other drugs being administered to the patient), theseverity of the particular disease undergoing therapy, and otherfactors, including the judgment of the prescribing medical practitioner.

Combination Therapy

The presently disclosed compounds may be used in combination with one ormore additional therapeutic agents that are used in the treatment,prevention, suppression or amelioration of the diseases or conditionsfor which compounds and compositions of the present invention areuseful. Such one or more additional therapeutic agents may beadministered, by a route and in an amount commonly used therefor,contemporaneously or sequentially with a compound or composition of thepresent invention. When a compound or composition of the presentinvention is used contemporaneously with one or more other drugs, apharmaceutical composition containing such other drugs in addition tothe compound or composition of the present invention is preferred.Accordingly, the pharmaceutical compositions of the present inventioninclude those that also contain one or more other active ingredients ortherapeutic agents, in addition to a compound or composition of thepresent invention.

Examples of the the one or more additional therapeutic agents arecorticosteroids, steroids, immunosuppressants, Immunoglobulin Gagonists, Dipeptidyl peptidase IV inhibitors, Lymphocyte functionantigen-3 receptor antagonists, Interleukin-2 ligands, Interleukin-1beta ligand inhibitors, IL-2 receptor alpha subunit inhibitors, HGF genestimulators, IL-6 antagonists, IL-5 antagonists, Alpha 1 antitrypsinstimulators, Cannabinoid receptor antagonists, Histone deacetylaseinhibitors, AKT protein kinase inhibitors, CD20 inhibitors, Abl tyrosinekinase inhibitors, JAK tyrosine kinase inhibitors, TNF alpha ligandinhibitors, Hemoglobin modulators, TNF antagonists, proteasomeinhibitors, CD3 modulators, Hsp 70 family inhibitors, Immunoglobulinagonists, CD30 antagonists, tubulin antagonists, Sphingosine-1-phosphatereceptor-1 agonists, connective tissue growth factor ligand inhibitors,caspase inhibitors, adrenocorticotrophic hormone ligands, Btk tyrosinekinase inhibitors, Complement Cls subcomponent inhibitors,Erythropoietin receptor agonists, B-lymphocyte stimulator ligandinhibitors, Cyclin-dependent kinase-2 inhibitors, P-selectinglycoprotein ligand-1 stimulators, mTOR inhibitors, Elongation factor 2inhibitors, Cell adhesion molecule inhibitors, Factor XIII agonists,Calcineurin inhibitors, Immunoglobulin G1 agonists, Inosinemonophosphate dehydrogenase inhibitors, Complement C₁s subcomponentinhibitors, Thymidine kinase modulators, Cytotoxic T-lymphocyteprotein-4 modulators, Angiotensin II receptor antagonists, AngiotensinII receptor modulators, TNF superfamily receptor 12A antagonists, CD52antagonists, Adenosine deaminase inhibitors, T-cell differentiationantigen CD6 inhibitors, FGF-7 ligands, dihydroorotate dehydrogenaseinhibitors, Syk tyrosine kinase inhibitors, Interferon type I receptorantagonists, Interferon alpha ligand inhibitors, Macrophage migrationinhibitory factor inhibitors, Integrin alpha-V/beta-6 antagonists,Cysteine protease stimulators, p38 MAP kinase inhibitors, TP53 geneinhibitors, Shiga like toxin I inhibitors, Fucosyltransferase 6stimulators, Interleukin 22 ligands, IRS1 gene inhibitors, Proteinkinase C stimulators, Protein kinase C alpha inhibitors, CD74antagonists, Immunoglobulin gamma Fc receptor IIB antagonists, T-cellantigen CD7 inhibitors, CD95 antagonists, N acetylmannosamine kinasestimulators, Cardiotrophin-1 ligands, Leukocyte elastase inhibitors,CD40 ligand receptor antagonists, CD40 ligand modulators, IL-17antagonists, TLR-2 antagonists, Mannan-binding lectin serine protease-2(MASP-2) inhibitors, Factor B inhibitors, Factor D inhibitors, C₃aRmodulators, C5aR² modulators, T cell receptor antagonists, PD-1inhibitors, PD-L1 inhibitors, TIGIT inhibitors, TIM-3 inhibitors, LAG-3inhibitors, VISTA inhibitors, STING agonists, IDO inhibitors, adenosinereceptor modulators, CD39 inhibitors, CD73 inhibitors, antagonists ofthe chemokine receptors, especially CXCR1, CXCR2, CXCR3, CXCR4, CXCR7,CCR1, CCR², CCR³, CCR4, CCR5, CCR7, CCR7, CCR9, CX3CR1 and CXCR6, andcombinations thereof.

In some embodiments, the additional therapeutic agent used in thetherapeutic methods herein, is selected from the group consisting ofobinutuzumab, rituximab, ocrelizumab, cyclophosphamide, prednisone,hydrocortisone, hydrocortisone acetate, cortisone acetate, tixocortolpivalate, prednisolone, methylprednisolone, triamcinolone acetonide,triamcinolone alcohol, mometasone, amcinonide, budesonide, desonide,fluocinonide, fluocinolone acetonide, halcinonide, betamethasone,betamethasone sodium phosphate, dexamethasone, dexamethasone sodiumphosphate, fluocortolone, hydrocortisone-17-valerate, halometasone,alclometasone dipropionate, beclomethasone, betamethasone valerate,betamethasone dipropionate, prednicarbate, clobetasone-17-butyrate,clobetasol-17-propionate, fluocortolone caproate, fluocortolonepivalate, fluprednidene acetate, hydrocortisone-17-butyrate,hydrocortisone-17-aceponate, hydrocortisone-17-buteprate, ciclesonideand prednicarbate, GB-0998, immuglo, begelomab, alefacept, aldesleukin,gevokizumab, daclizumab, basiliximab, inolimomab, beperminogeneperplasmid, sirukumab, tocilizumab, clazakizumab, mepolizumab,fingolimod, panobinostat, triciribine, nilotinib, imatinib, tofacitinib,momelotinib, peficitinib, itacitinib, infliximab, PEG-bHb-CO,etanercept, ixazomib, bortezomib, muromonab, otelixizumab, gusperimus,brentuximab vedotin, Ponesimod, KRP-203, FG-3019, emricasan,corticotropin, ibrutinib, cinryze, conestat, methoxy polyethyleneglycol-epoetin beta, belimumab, blisibimod, atacicept, seliciclib,neihulizumab, everolimus, sirolimus, denileukin diftitox, LMB-2,natalizumab, catridecacog, ciclosporin, tacrolimus, voclosporin,voclosporin, canakinumab, mycophenolate, mizoribine, CE-1145, TK-DLI,abatacept, belatacept, olmesartan medoxomil, sparsentan, TXA-127,BIIB-023, alemtuzumab, pentostatin, itolizumab, palifermin, leflunomide,

PRO-140, cenicriviroc, fostamatinib, anifrolumab, sifalimumab, BAX-069,BG-00011, losmapimod, QPI-1002, ShigamAbs, TZ-101, F-652, reparixin,ladarixin, PTX-9908, aganirsen, APH-703, sotrastaurin, sotrastaurin,milatuzumab, SM-101, T-Guard, APG-101, DEX-M74, cardiotrophin-1,tiprelestat, ASKP-1240, BMS-986004, HPH-116, KD-025, OPN-305, TOL-101,defibrotide, pomalidomide, Thymoglobulin, laquinimod, remestemcel-L,Equine antithymocyte immunoglobulin, Stempeucel, LIV-Gamma, Octagam 10%,t2c-001, 99mTc-sestamibi, Clairyg, Prosorba, pomalidomide, laquinimod,teplizumab, FCRx, solnatide, foralumab, ATIR-101, BPX-501, ACP-01,ALLO-ASC-DFU, irbesartan+propagermanium, ApoCell, cannabidiol, RGI-2001,saratin, anti—CD3 bivalent antibody-diphtheria toxin conjugate, NOX-100,LT-1951, OMS721, ALN—CCS, ACH-4471, AMY-101, Acthar gel, andCD4+CD25+regulatory T-cells, MEDI7814, P32, P59, pembrolizumab,nivolumab, atezolizumab, avelumab, durvalumab, CCX354, CCX721, CCX9588,CCX140, CCX872, CCX598, CCX6239, CCX587, CCX624, CCX282, CCX025, CCX507,CCX430, CCX765, CCX758, CCX771, CCX662, CCX650, and combinationsthereof.

The disease or disorder being treated will determine which additionaltherapeutic agent or therapeutic agents are most appropriatelyadministered in combination with the compounds of the presentinvention—such determination can be made by a person of skill in theart.

The weight ratio of the compound of the present invention to the secondactive ingredient may be varied and will depend upon the effective doseof each ingredient. Generally, an effective dose of each will be used.Thus, for example, when a compound of the present invention is combinedwith an NSAID the weight ratio of the compound of the present inventionto the NSAID will generally range from about 1000:1 to about 1:1000,preferably about 200:1 to about 1:200. Combinations of a compound of thepresent invention and other active ingredients will generally also bewithin the aforementioned range, but in each case, an effective dose ofeach active ingredient should be used.

Non-Pharmaceutical Applications

In another aspect of the invention, the compounds of the invention canbe used in a variety of non-pharmaceutical in vitro and in vivoapplication. For example, the compounds of the invention may be labeledand used as probes for the detection and localization of C5a receptor(cell preparations or tissue sections samples). The compounds of theinvention may also be used as positive controls in assays for C5areceptor activity, i.e., as standards for determining the ability of acandidate agent to bind to C5a receptor, or as radiotracers for positronemission tomography (PET) imaging or for single photon emissioncomputerized tomography (SPECT). Such methods can be used tocharacterize C5a receptors in living subjects. For example, a C5areceptor modulator may be labeled using any of a variety of well knowntechniques (e.g., radiolabeled with a radionuclide such as tritium), andincubated with a sample for a suitable incubation time (e.g., determinedby first assaying a time course of binding). Following incubation,unbound compound is removed (e.g., by washing), and bound compounddetected using any method suitable for the label employed (e.g.,autoradiography or scintillation counting for radiolabeled compounds;spectroscopic methods may be used to detect luminescent groups andfluorescent groups). As a control, a matched sample containing labeledcompound and a greater (e.g., 10-fold greater) amount of unlabeledcompound may be processed in the same manner. A greater amount ofdetectable label remaining in the test sample than in the controlindicates the presence of C5a receptor in the sample. Detection assays,including receptor autoradiography (receptor mapping) of C5a receptor incultured cells or tissue samples may be performed as described by Kuharin sections 8.1.1 to 8.1.9 of Current Protocols in Pharmacology (1998)John Wiley & Sons, New York.

The compounds provided herein may also be used within a variety of wellknown cell separation methods. For example, modulators may be linked tothe interior surface of a tissue culture plate or other support, for useas affinity ligands for immobilizing and thereby isolating,

C5a receptors (e.g., isolating receptor-expressing cells) in vitro. Inone preferred application, a modulator linked to a fluorescent marker,such as fluorescein, is contacted with the cells, which are thenanalyzed (or isolated) by fluorescence activated cell sorting (FACS).

I. EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR spectra wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In the examples, a single m/e value isreported for the M+H (or, as noted, M−H) ion containing the most commonatomic isotopes. Isotope patterns correspond to the expected formula inall cases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP1100 HPLC for sample delivery. Normally the analyte was dissolvedin methanol at 0.1 mg/mL and 1 microliter was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

The following abbreviations are used in the Examples and throughout thedescription of the invention:

-   EtOH: Ethanol-   EtONa: Sodium ethoxide-   THF: Tetrahydrofuran-   TLC: Thin layer chromatography-   MeOH: Methanol

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. One skilled in the art will also recognize that alternativemethods may be employed to synthesize the target compounds of thisinvention, and that the approaches described within the body of thisdocument are not exhaustive, but do provide broadly applicable andpractical routes to compounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

Example 1 Synthesis of1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

Step a: A mixture of 3-methyl-2-nitro-phenol (50.0 g, 326.5 mmol),1-iodo-2-methyl-propane (184.0 g, 1.0 mol) and Cs₂CO₃ (326.0 g, 1.0 mol)in acetone (500 mL) was stirred overnight under reflux. It was thencooled to room temeperature and filtered through Celite.

The filtrate was collected and concentrated under reduced pressure. Theobtained solid was dissolved into EtOAc, washed with brine, dried overNa₂SO₄ and concentrated under reduced pressure to afford1-isobutoxy-3-methyl-2-nitro-benzene. ¹H NMR (400 MHz, CDCl₃) δ 7.26 (t,J=8.0 Hz, 1H), 6.82 (m, 2H), 3.78 (d, J=6.8 Hz, 2H), 2.94 (s, 3H), 2.07(m, 1H), 0.98 (d, J=6.4 Hz, 6H).

A pressure vessel containing 1-isobutoxy-3-methyl-2-nitro-benzene (130.4g, 623.2 mmol), 10% Pd/C (25 g, 50% wet) and EtOH (750 mL) was agitatedunder a hydrogen atmosphere at 45 psi for 3 h. The mixture was filteredthrough Celite. The filtrate was collected and concentrated underreduced pressure to yield 2-isobutoxy-6-methyl-aniline. C₁₁H₁₈NO [M+H]⁺180.2, found 180.2.

Caution: Diazonium Formation could be Potentially Dangerous, PleaseHandle with Care and Ware Proper Personal Protection Equipment!

Step b: To 100 mL of concentrated HCl at −10° C. was addedisobutoxy-6-methyl aniline (26.4 g, 147.3 mmol) portionwise to obtain astirrable suspension. After stirring for 30 min at the same temperature,a solution of NaNO₂ (12.2 g, 176.8 mmol) in water (25 mL) was addeddropwise over 20 min to obtain the diazonium salt.

To the above diazonium salt was added tin(II) chloride dihydrate (83.0g, 367.8 mmol) in concentrated HCl (120 mL) portionwise. The obtainedmixture was then stirred for 10 min at −10° C. followed by 1 h at roomtemperature. The mixture was then diluted into DCM (400 mL) and water.The organic layer was separated, dried over Na₂SO₄ and concentrated on arotary evaporator under reduced pressure to yield(2-isobutoxy-6-methyl-phenyl)hydrazine hydrochloride. C₁₁H₁₉N₂O [M+H]⁺195.1, found 195.1.

Step c: To a stirred suspension of (2-isobutoxy-6-methylphenyl)hydrazinehydrochloride (8.0 g, 39.9 mmol) in EtOH (60 mL) and glacial acetic acid(12 mL, 208 mmol) was added tert-butyl3-cyano-4-oxopiperidine-1-carboxylate (5.0 g, 22.3 mmol) at roomtemperature. The resulting mixture was stirred under reflux for 16 h.After removal of solvent under reduced pressure, the residue wasdissolved in EtOAc and washed with 2 N aqueous NaOH, brine, and driedover MgSO₄. The solvent was removed under reduced pressure and theresidue was purified by silica gel flash chromatography (5 to 55% EtOAcin hexanes) to give tert-butyl3-amino-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₂H₃₃N₄O₃ [M+H]⁺ 401.2, found 401.2.

Caution: Diazonium Formation could be Potentially Dangerous, PleaseHandle with Care and Ware Proper Personal Protection Equipment!

Isoamyl nitrite (96%, 4.0 mL, 28.6 mmol) was added slowly at roomtemperature to a mixture of tert-butyl3-amino-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(3.0 g, 8.1 mmol), CuBr (4.0 g, 27.9 mmol) and MeCN (50 mL) in a 250 mLround bottom flask under magnetic stirring. The resulting mixture wasstirred at room temperature for 1 h, diluted with EtOAc, filteredthrough Celite, washed with saturated aqueous NH₄Cl solution, and driedover MgSO₄. The solvent was removed under reduced pressure and theresidue was purified by silica gel flash chromatography (2 to 25% EtOAcin hexanes) to give tert-butyl3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₂H₃₁BrN₃O₃ [M+H]⁺ 464.1, found 464.2.

Step d: A mixture of 4-bromo-2,5-difluoroaniline (1.5 g, 7.2 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (2.2 g, 8.7mmol), KOAc (1.8 g, 18.3 mmol) and Pd(dppf)Cl₂ complex withdichloromethane (580.0 mg, 0.7 mmol) in dioxane (12 mL) was stirred at95° C. for 2 h under nitrogen. The mixture was then cooled to roomtemperature and filtered over Celite. The filtrate was collected,concentrated under reduced pressure, and purified by silica gel flashchromatography (0 to 50% EtOAc in hexanes) to give2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline. MS:(ES) m/z calculated for C₁₂H₁₇BF₂NO₂ [M+H]⁺ 256.1, found 256.2.

To a suspension of tert-butyl3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(125.0 mg, 0.3 mmol),2,5-difluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(75.0 mg, 0.3 mmol), Na₂CO₃ (85.0 mg, 0.8 mmol) in dioxane (4 mL) andwater (1 mL) was added Pd(dppf)Cl₂ complex with dichloromethane (26.0mg, 0.03 mmol). The reaction mixture was degassed (N₂) for 2 min andstirred under N₂ at 95° C. for 6 h. The reaction mixture was dilutedwith EtOAc, filtered through Celite, washed with brine, and dried overMgSO₄. The solvent was removed under reduced presure and the residue waspurified by silica gel flash chromatography (5 to 40% EtOAc in hexanes)to give tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₈H₃₅F₂N₄O₃ [M+H]⁺ 513.3, found 513.3.

Step e: To a stirred solution of tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(0.5 g, 2.0 mmol) in anhydrous THF (5 mL) was addedN,N-disopropylethylamine (0.6 g, 4.8 mmol) and trimethylsilylisocyanate(0.3 g, 2.6 mmol). The reaction mixture was stirred at room temperaturefor 16 h. After completion, the reaction mixture was diluted with EtOAc,washed with brine, and dried over MgSO₄. The solvent was removed underreduced presure and the residue was purified by silica gel flashchromatography (5 to 40% EtOAc in hexanes) to affordtert-butyl-3-(2,5-difluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated C₂₉H₃₆F₂N₅O₄ [M+H]⁺ 556.3, found 556.3.

Step f: To a solution oftert-butyl-3-(2,5-difluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.2 g, 3.6 mmol), in dichloromethane (10 mL) was added 4 N HCl indioxane (3.0 mL, 12.0 mmol). The resulting mixture was stirred at roomtemperature for 2 h. After completion of the reaction, the solvent wasdiluted with water and saturated aqueous NaHCO₃ and extracted withEtOAc, washed with brine and dried over MgSO₄. The solvent was removedunder reduced presure to give1-(2,5-difluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea.MS: (ES) m/z calculated for C₂₄H₂₈F₂N₅O₂ [M+H]⁺ 456.2, found 456.2.

Step g: To a suspension of1-(2,5-difluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea(3.0 g, 6.6 mmol) in DMSO (10 mL) was added 3,5-dichloro-5-fluoropyridine (1.6 g, 9.9 mmol) and Li₂CO₃ (1.9 g, 25.7 mmol) at roomtemperature. The resulting mixture was stirred at 90° C. for 4 h. Aftercompletion of the reaction, the mixture was cooled to room temperature,and diluted with EtOAc. The organic layer was washed with brine, anddried over MgSO₄. The solvent was removed under reduced pressure and theresidue was purified by silica gel flash chromatography (10 to 60% EtOAcin hexanes) to afford1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea.¹HNMR (400 MHz, CD₃OD): δ 8.15 (d, J=2.3 Hz, 1H), 8.02 (dd, J=7.1, 12.5Hz, 1H), 7.84 (d, J=2.3 Hz, 1H), 7.32 (t, J=7.8 Hz, 1H), 6.92 (d, J=8.2Hz, 1H), 6.89 (d, J=7.5 Hz, 1H), 6.82 (dd, J=6.3, 11.3 Hz, 1H), 4.87(br, 2H), 4.39 (s, 2H), 3.70- 3.82 (m, 4H), 3.65 (dd, J=7.1, 9.0 Hz,1H), 3.07 (t, J=6.7 Hz, 2H), 2.02 (s, 3H), 1.85-1.95 (m, 1H), 0.85 (d,J=7.0 Hz, 6H). MS: (ES) m/z calculated C₂₉H₂₉Cl₂F₂N₆O₂ [M+H]⁺ 601.2,found 601.5.

Example 2 Synthesis of4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzamide

Step a: A mixture of tert-butyl3-bromo-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(120.0 mg, 0.26 mmol), (4-cyanophenyl)boronic acid (100.0 mg, 0.68mmol), Pd(PPh₃)₄ (45.0 mg, 0.04 mmol) and K₂CO₃ (125.0 mg, 0.9 mmol) intoluene (2 mL) and water (0.3 mL) was stirred at 110° C. for 3 h underN₂. The mixture was cooled to room temperature, quenched with water, andextracted with EtOAc. The organic layer was separated, washed withbrine, dried over Na₂SO₄, and filtered. The solvent was concentratedunder reduced pressure and the residue was purified by silica gel flashchromatography (0 to 40% EtOAc in hexanes) to yield tert-butyl3-(4-cyanophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₉H₃₅N₄O₃ [M+H]⁺ 487.3, found 487.2.

Step b: A mixture of tert-butyl3-(4-cyanophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(92.0 mg, 0.2 mmol) and 4 N HCl in dioxane (2.0 mL, 8.0 mmol) indichloromethane (2 mL) was stirred at room temperature for 1 h. Themixture was basified with saturated aqueous NaHCO₃ and extracted withEtOAc. The organic layer was separated, washed with brine, dried overNa₂SO₄, and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatograph(0 to 25% MeOH in dichloromethane) to yield4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile.MS: (ES) m/z calculated for C₂₄H₂₇N₄O [M+H]⁺ 387.2, found 387.2.

Step c: A mixture of4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile(56.0 mg, 0.15 mmol), 2-bromo-4-(tert-butyl)-1-methylbenzene (131.0 mg,0.58 mmol), Pd(OAc)₂ (12.0 mg, 0.05 mmol), X-Phos (60.0 mg, 0.13 mmol)and Cs₂CO₃ (141.0 mg, 0.43 mmol) in dioxane (2 mL) was stirred at 110°C. for 1 h under N₂. The mixture was cooled to room temperature, dilutedwith water, and extracted with EtOAc. The organic layer was separated,washed with brine, dried over Na₂SO₄, and filtered. The solvent wasconcentrated under reduced pressure and the residue was purified bysilica gel flash chromatography (0 to 35% EtOAc in hexanes) to yield4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile.MS: (ES) m/z calculated for C₃₅H₄₁N₄O [M+H]⁺ 533.3, found 533.3.

Step d: To a mixture of4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzonitrile(36.0 mg, 0.07 mmol) in DCM (1 mL) and DMSO (6 mL) was added 4 N aqueousNaOH (1.0 mL, 4.0 mmol) and H₂O₂ (0.40 mL, 35% in water). The mixturewas stirred for 30 min at room temperature, diluted with water, andextracted with EtOAc. The organic layer was separated, washed withbrine, dried over Na₂SO₄, and filtered. The solvent was concentratedunder reduced pressure and the residue was purified by silica gel flashchromatography (0 to 65% EtOAc in hexanes) to yield4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(24sobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)benzamide.¹H NMR (400 MHz, CDCl₃): 8 7.65 (dd, J=8.0, 8.4 Hz, 2H), 7.12-7.24 (m,5H), 7.05 (dd, J=8.0, 2.0 Hz, 1H), 6.81 (d, J=7.2 Hz, 1H), 6.70 (d,J=8.4 Hz, 1H), 6.05 (br s, 1H), 5.73 (br s, 1H) 4.11 (m, 2H), 3.56 (m,2H), 3.36 (m, 2H), 3.01 (m, 2H), 2.31 (s, 3H), 2.09 (s, 3H), 1.86 (m,1H), 1.31 (s, 9H), 0.81 (m, 6H). MS: (ES) m/z calculated for C₃₅H₄₃N₄O₂[M+H]⁺ 551.3, found 551.3.

Example 3 Synthesis of1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoropheny)urea

Step a: To a mixture of 2,5-difluoro-4-nitrobenzoic acid (35.5 g, 174.8mmol) in dichloromethane (400 mL) at room temperature, was added slowlyoxalyl chloride (16.0 mL, 186.3 mmol), followed by the addition of DMF(0.2 mL, 2.6 mmol). The resulting mixture was stirred at roomtemperature overnight. It was then concentrated in vacuo to yield2,5-difluoro-4-nitrobenzoyl chloride and used directly for next step.

Step b: To a mixture of tert-butyl 4-oxopiperidine-1-carboxylate (35.6g, 175.8mmo1) in dichloromethane (500 mL) at 0° C. was added MgCl₂ (34.0g, 357.1 mmol), 2,5-difluoro-4-nitrobenzoyl chloride (38.8 g, 175.1mmol) and triethylamine (50.0 mL, 355.8 mmol) sequentially. The mixturewas stirred for 30 min at 0° C. followed by 7 h at room temperature. Itwas then cooled to 0° C., quenched with saturated aqueous NH₄Cl andextracted with dichloromethane. The organic layer was separated, washedwith brine, dried over Na₂SO₄ and filtered. The solvent was concentratedunder reduced pressure to afford tert-butyl3-(2,5-difluoro-4-nitrobenzoyl)-4-oxopiperidine-1-carboxylate. MS: (ES)m/z calculated for C₁₇H₁₉F₂N₂O₆Na [M+Na]⁺ 385.1, found 385.1.

Step c: A mixture of tert-butyl3-(2,5-difluoro-4-nitrobenzoyl)-4-oxopiperidine-1-carboxylate (30.0 g,78.1 mmol) and (2,6-dimethylphenyl)hydrazine hydrochloride (17.5 g,101.5 mmol) in EtOH (30 mL) was heated to 75° C. for 4 h. The mixturewas then cooled to room temperature, concentrated under reduced pressureand the residue was purified by silica gel flash chromatography (0 to30% EtOAc in hexanes) to give tert-butyl3-(2,5-difluoro-4-nitrophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₅H₂₆F₂N₄O₄ [M+H]⁺ 485.2, found 485.2.

Step d: A mixture of tert-butyl3-(2,5-difluoro-4-nitrophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(10.0 g, 20.6 mmol) and PtO₂ (0.5 g, 2.2 mmol) in EtOAc (100 mL) wasagitated in a Parr shaker bottle under hydrogen at 40 psi overnight. Themixture was filtered through Celite and concentrated in vacuo to affordtert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₅H₂₈F₂N₄O₂ [M+H]⁺ 455.2, found 455.2.

Step e: A mixture of3-(4-amino-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(7.9 g, 17.3 mmol) and benzoyl isocyanate (2.6 g, 17.3 mmol) in THF (40mL) was stirred for 3 h at room temperature. The mixture wasconcentrated under reduced pressure to obtain tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

A mixture of tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(˜17.3 mmol, from above) and K₂CO₃ (7.1 g, 51.3 mmol) in MeOH (100 mL)was stirred for 2 h at room temperature followed by 20 min at 50° C. Themixture was extracted with IPA:CHCl₃ (1:3). The organic layer wasseparated, dried over Na₂SO₄, concentrated under reduced pressure andpurified by silica gel flash chromatography (0 to 70% EtOAc in hexanes)to afford tert-butyl3-(2,5-difluoro-4-ureidophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₆H₂₉F₂N₅O₃ [M+H]⁺ 498.2, found 498.2

Step f: A mixture of tert-butyl3-(2,5-difluoro-4-ureidophenyl)-2-(2,6-dimethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(7.0 g, 14.1 mmol) and 4 N HCl in dioxane (14.0 mL, 56.0 mmol) indichloromethane (20 mL) was stirred at room temperature overnight. Themixture was then concentrated in vacuo to yield1-(4-(2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride. MS: (ES) m/z calculated for C₂₁H₂₁F₂N5O [M+H]⁺ 398.2,found 398.2.

Step g: Triethylamine (3.9 mL, 27.7 mmol) was added to a suspension of1-(4-(2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride (4 g, 9.23 mmol), 3,5-dichloro-2-fluoropyridine (1.7 g,10.2 mmol), and Li₂CO₃ (2.0 g, 27.7 mmol) in DMSO (14 mL) under magneticstirring. The resulting mixture was stirred at 90° C. for 4 h. Aftercooling to room temperature, the reaction mixture was diluted with DCM,washed with brine, and dried over MgSO₄. The solvent was removed underreduced pressure and the residue was purified by silica gel flashchromatography (20% to 50% EtOAc in hexanes, followed by 0% to 50% EtOAcin dichloromethane) followed by recrystallization inMeOH/dichloromethane/EtOAc to afford1-(4-(5-(3,5-dichloropyridin-2-yl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea.¹H NMR (400 MHz, CD₃OD): δ 8.16 (d, J=2.3 Hz, 1H), 8.01 (dd, J=6.6, 12.5Hz, 1H), 7.83 (d, J=2.3 Hz, 1H), 7.25 (dd, J=7.0, 8.2 Hz, 1H), 7.06-7.16(m, 2H), 6.70 (dd, J=6.6, 11.4 Hz, 1H), 4.86 (br, 3H), 4.37 (s, 2H),3.76 (t, J=5.8 Hz, 2H), 3.03 (t, J=5.8 Hz, 2H), 1.99 (s, 6H). MS: (ES)m/z calculated C₂₆H₂₃C₁₂F₂N₆O [M+H]⁺ 543.1, found 543.1.

Example 4 Synthesis of1-(4-(2-(2,6-diethylphenyl)-5-(tert-pentyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

Caution: Diazonium Formation could be Potentially Dangerous, PleaseHandle with Care and Wear Proper Personal Protection Equipment!

Step a: To a 250 mL flask charged with 90 mL of concentratedhydrochloric acid under magnetic stirring was added 2,6-diethylaniline(10.0 g, 67 mmol). The resulting mixture was stirred for 30 min andcooled with an ice-salt bath until the internal temperature reached −5°C. A solution of sodium nitrite (5.5 g, 80 mmol) in water (60 mL) wasadded slowly to the above mixture while maintaining the internaltemperature below 5° C.

Separately, tin(II) chloride dihydrate (31.6 g, 140 mmol) was added to a500 mL 3-neck round bottom flask charged with concentrated hydrochloricacid (60 mL) under mechanical stirring. The resulting solution was thencooled with an ice bath.

The diazonium slurry was then filtered into the 500 mL flask containingthe cooled tin chloride solution with vigorous stirring. After 90 min,the reaction mixture was transferred to a 500 mL Erlenmeyer flask andthe flask was rinsed with water (20 mL) and chloroform (8 mL). Thecombined mixture was stirred overnight at room temperature. The entireliquid layer was decanted to give a wet solid. The recovered materialwas dried in vacuo for one day and then transferred to a 500 mL 3-neckround bottom flask equipped with an overhead mechanical stirrer andstirred with ether (180 mL). The resulting mixture was cooled in an icebath, and NaOH solution (10 N, 30 mL) was added slowly to the abovemixture while maintaining the inner temperature below 12° C. After theaddition, the mixture was allowed to stand for 2 h on ice. The etherlayer was decanted into a 500 mL flask and a stream of hydrogen chloridegas was bubbled into the ether solution while stirring. The resultingprecipitate was collected by filtration to afford(2,6-diethylphenyl)hydrazine hydrochloride. MS: (ES) m/z calculated forC₁₀H₁₇N₂ [M+H]⁺ 165.1, found 165.1.

Step b: A mixture of tert-butyl3-(2,5-difluoro-4-nitrobenzoyl)-4-oxopiperidine-1-carboxylate (60.0 g,156.1 mmol) and (2,6-diethylphenyl)hydrazine hydrochloride (30.0 g,149.5 mmol) in EtOH (560 mL) was heated to 50° C. for 3 h. The mixturewas then cooled to room temperature and quenched with saturated aqueousNaHCO₃. The mixture was extracted with EtOAc, the organic layer wasseparated, washed with brine, dried over Na₂SO₄ and filitered. Thesolvent was concentrated under reduced pressure and the residue waspurified by silica gel flash chromatography (0 to 50% EtOAc in hexanes)to afford tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo [4,3-c]pyridine-5-carboxylate. MS: (ES) m/z calculated forC₂₇H₃₁F₂N₄O₄ [M+H]⁺ 513.2, found 513.5.

Step c: A mixture of tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(9.6 g, 18.7 mmol), iron powder (10.0 g, 179.1 mmol) and NH₄Cl (15.0 g,280.4 mmol) in EtOH (200 mL) and water (20 mL) was heated to 80° C. for1 h. The mixture was then cooled to room temperature, filtered overCelite and washed with EtOAc. The filtrate was partitioned between EtOAcand saturated aqueous NaHCO₃. The organic layer was separated, driedover Na₂SO₄ and filtered. The solvent was concentrated in vacuo toafford tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₇H₃₃F₂N₄O₂ [M+H]⁺ 483.3, found 483.3.

Step d: A mixture of tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(9.2 g, 19.1 mmol) and benzoyl isocyanate (11.3 g, 76.8 mmol) in THF(100 mL) was stirred at room temperature for 4 h. The mixture was thenconcentrated in vacuo to afford tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

A mixture of tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(-19.1 mmol) from above and K₂CO₃ (10.0 g, 72.3 mmol) in MeOH (100 mL)was stirred for 7 h at room temperature. The reaction mixture wasextracted with IPA:CHCl₃ (1:3). The organic layer was separated, driedover Na₂SO₄ and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatography(0 to 100% EtOAc in dichloromethane follwed by 0 to 20% MeOH indichloromethane) to afford tert-butyldiethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₈H₃₄F₂N₅O₃ [M+H]⁺ 526.3, found 526.3.

Step e: A mixture of tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(2.9 g, 5.5 mmol) and 4 N HCl in dioxane (35.0 mL, 140.0 mmol) indichloromethane (30 mL) was stirred at room temperature for 1.5 h. Themixture was then concentrated in vacuo to yield1-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride. MS: (ES) m/z calculated for C₂₃H₂₆F₂N₅O [M+H]⁺ 426.2,found 426.2.

Step f: To a mixture of1-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride (115.0 mg, 0.3 mmol) in THF (2.5 mL) at 0° C. was added3-chloro-3-methylbut-1-yne (32 mg, 0.3 mmol), NEt₃ (88 μL, 0.6 mmol) andCuCl (30 mg, 0.3 mmol) sequentially. The mixture was allowed to warm toroom temperature and stirred for 30 min. The mixture was then basifiedwith saturated aqueous NaHCO₃ and extracted with EtOAc. The organiclayer was separated, washed with brine, dried over Na₂SO₄, and filtered.The solvent was concentrated under reduced pressure and the residue waspurified by silica gel flash chromatography (0 to 100% EtOAc in hexanes)to afford1-(4-(2-(2,6-diethylphenyl)-5-(2-methylbut-3-yn-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea.MS: (ES) m/z calculated for C₂₈₁H₃₂F₂N₅O [M+H]⁺ 492.3, found 492.2.

Step g: A mixture of1-(4-(2-(2,6-diethylphenyl)-5-(2-methylbut-3-yn-2-yl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea(66.0 mg, 0.13 mmol) and Pd/C (50 mg, 50% in water) in EtOAc (35 mL) wasagitated in a Parr shaker bottle under hydrogen at 53 psi for 1.5 h. Themixture was filtered over Celite. The filtrate was concentrated underreduced pressure and the residue was purified by HPLC (MeCN/H₂O, with0.1% TFA), basified with saturated aqueous NaHCO₃ and extracted withEtOAc. The organic layer was separated, washed with brine, dried overNa₂SO₄, filtered, and treated with 1.0 M HCl in dietheyl ether. Thesolvent was concentrated in vacuo to yield1-(4-(2-(2,6-diethylphenyl)-5-(tert-pentyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureaas hydrochloride salt. ¹H NMR (400 MHz, CD₃OD): δ 8.15 (br s, 1H), 7.47(dd, J=7.4, 7.4 Hz, 1H), 7.29 (m, 2H), 6.68 (br s, 1H), 4.57 (br s, 2H),4.18 (br s, 1H), 3.62 (br s, 1H), 3.32 (br s, 2H), 2.38 (br s, 2H), 2.22(br s, 2H), 1.98 (br s, 2H), 1.70 (s, 1H), 1.55 (br s, 6H), 1.55 (br s,6H), 1.32-1.42 (m, 3H), 1.00-1.22 (m, 9H). MS: (ES) m/z calculated forC₂₈H₃₆F₂N₅O [M+H]⁺ 496.3, found 496.3.

Example 5 Synthesis ofN-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)formamide

Step a: A mixture of tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.7 g, 3.5 mmol), acetyl chloride (1.4 mL, 19.6 mmol) and NEt₃ (3.7 mL,26.1 mmol) in THF (120 mL) was stirred at room temperature for 2 h. Thereaction mixture was quenched with saturated aqueous NaHCO₃ andextracted with EtOAc. The organic layer was separated, washed withbrine, dried over Na₂SO₄ and filtered. The solvent was concentratedunder reduced pressure and the residue was purified by silica gel flashchromatography (0 to 80% EtOAc in hexanes) to afford tert-butyl3-(4-acetamido-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₉H₃₅F₂N₄O₃ [M+H]⁺ 525.3, found 525.6.

Step b: A mixture of tert-butyl3-(4-acetamido-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.3 g, 2.4 mmol) and 4 N HCl in dioxane (20.0 mL, 80.0 mmol) indichloromethane (20 mL) was stirred at room temperature for 0.5 h. Themixture was then concentrated in vacuo to yieldN-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamidehydrochloride. MS: (ES) m/z calculated for C₂₄H₂₇F₂N₄O [M+H]⁺ 425.2,found 425.2.

Step c: A mixture ofN-(4-(2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamidehydrochloride (0.45 g, 1.0 mmol), 2,4-bis(trifluoromethyl)benzaldehyde(0.7 g, 2.9 mmol), NaBH(OAc)₃ (0.8 g, 3.8 mmol), NEt₃ (0.5 mL, 3.6 mmol)and HOAc (0.2 mL, 3.3 mmol) in dichloromethane (5 mL) was stirred at 30°C. for 1 h. The reaction was quenched with saturated aqueous NaHCO₃ andextracted with EtOAc. The organic layer was separated, washed withbrine, dried over Na₂SO₄, and filtered. The solvent was concentratedunder reduced pressure and the residue was purified by silica gel flashchromatography (0 to 80% EtOAc in hexanes) to affordN-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide.MS: (ES) m/z calculated for C₃₃H₃₀F₈N₄O [M+H]⁺ 651.3, found 651.6.

Step d: A mixture ofN-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)acetamide(0.5 g, 0.8 mmol) and 4 N HCl in dioxane (3.0 mL, 12.0 mmol) in water(0.6 mL) was stirred at 80° C. for 40 min. The mixture was cooled toroom temperature, basified with saturated aqueous NaHCO₃ and extractedwith EtOAc. The organic layer was separated, washed with brine, driedover Na₂SO₄, and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatography(0 to 70% EtOAc in hexanes) to afford4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoroaniline.MS: (ES) m/z calculated for C₃₁H₂₉F₈N₄ [M+H]⁺ 609.2, found 609.2.

Step e: A mixture of4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoroaniline(25.0 mg, 0.04 mmol) and HCO₂H (1 mL) was stirred at 75° C. for 40 min.The mixture was cooled to room temperature, basified with saturatedaqueous NaHCO₃ and extracted with EtOAc. The organic layer wasseparated, washed with brine, dried over Na₂SO₄, and filtered. Thesolvent was concentrated under reduced pressure and the residue waspurified by silica gel flash chromatography (0 to 70% EtOAc in hexanes)to affordN-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)formamide.¹H NMR (400 MHz, CDCl₃): δ 8.38 (d, J=1.2 Hz, 1H), 8.15 (m, 2H), 7.89(br s, 1H), 7.79 (d, J=8.4 Hz, 1H), 7.53 (br s, 1H), 7.28 (dd, J=7.6,7.6 Hz, 1H), 7.10 (d, J=7.6 Hz, 2H), 6.55 (m, 1H), 3.95 (s, br, 2H),3.58 (bs, 2H), 2.93 (m, 4H), 2.31 (sextet, J=7.6 Hz, 2H), 2.19 (sextet,J=7.2 Hz, 2H), 1.08 (t, J=7.4 Hz, 6H). MS: (ES) m/z calculated forC₃₂H₂₉F₈N₄O [M+H]⁺ 637.2, found 637.2.

Example 6 Synthesis of1-(4-(2-(2,6-diethylphenyl)-5-(2-hydroxy-2-methyl-1-phenylpropyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

Step a: A mixture of tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.0 g, 2.0 mmol) and 4 N HCl in dioxane (5.0 mL, 20.0 mmol) indichloromethane (10 mL) was stirred at room temperature for 1.5 h. Themixture was then concentrated in vacuo to yield2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridinehydrochloride. MS: (ES) m/z calculated for C₂₂H₂₃F₂N₄O₂ [M+H]⁺ 413.2,found 413.2.

N,N-diisopropylethylamine (3.0 mL, 17.3 mmol) was added to a suspensionof the above2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridinehydrochloride (˜2.0 mmol), and ethyl 2-bromo-2-phenylacetate (2.0 mL,11.4 mmol) in acetonitrile (6 mL) under magnetic stirring. The resultingmixture was stirred at 90° C. for 3 h. After cooling to roomtemperature, the reaction mixture was diluted with EtOAc, washed withbrine and dried over MgSO₄. The solvent was removed under reducedpressure and the residue was purified by silica gel flash chromatography(10% to 30% EtOAc in hexanes) to afford ethyl2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate.MS: (ES) m/z calculated for C₃₂H₃₃F₂N₄O₄ [M+Na]⁺ 575.2, found 575.3.

Step b: A mixture of ethyl2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate(1.0 g, 1.7 mmol), iron powder (1.0 g, 17.9 mmol) and NH₄Cl (1.5 g, 26.9mmol) in EtOH (20 mL) and water (2 mL) was heated to 80° C. for 1 h. Themixture was then cooled to room temperature, filtered over Celite andwashed with EtOAc. The filtrate was partitioned between EtOAc andsaturated aqueous NaHCO₃. The organic layer was separated, dried overNa₂SO₄, and filtered. The solvent was concentrated in vacuo to affordethyl2-(3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate.MS: (ES) m/z calculated for C₃₂H₃₅F₂N₄O₂ [M+H]⁺ 545.3, found 545.3.

Step c: A mixture of ethyl2-(3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate(800.0 mg, 1.5 mmol) and benzoyl isocyanate (1.1 g, 7.7 mmol) in THF (10mL) was stirred at room temperature for 4 h. The mixture was thenconcentrated in vacuo to afford ethyl2-(3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate.

A mixture of ethyl2-(3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate(˜1.5 mmol) from above and K₂CO₃ (1.0 g, 7.2 mmol) in MeOH (10 mL) wasstirred at room temperature overnight. The reaction mixture wasextracted with EtOAc. The organic layer was separated, dried over Na₂SO₄and filtered. The solvent was concentrated under reduced pressure andthe residue was purified by silica gel flash chromatography (10 to 45%EtOAc in hexanes) to afford ethyl2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate.MS: (ES) m/z calculated for C₃₃H₃₆F₂N₅O₃ [M+H]⁺ 588.3, found 588.3.

Step d: To a solution of ethyl2-(2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridin-5-yl)-2-phenylacetate(100.0 mg, 0.2 mmol) in THF (5 mL) was added 1.6 M CH₃Li solution indiethyl ether (0.7 mL, 1.1 mmol) at 0° C. The obtained mixture wasstirred at the same temperature for 30 min, quenched with saturatedNH₄Cl and extracted with EtOAc. The organic layer was separated, washedwith brine, dried over Na₂SO₄, and filtered. The solvent was removedunder reduced pressure and the residue was purified by preparative TLC(40% EtOAc in hexanes followed by 30% EtOAc in dichloromethane) toafford1-(4-(2-(2,6-diethylphenyl)-5-(2-hydroxy-2-methyl-1-phenylpropyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea.¹H NMR (400 MHz, CD₃OD): δ 7.93 (dd, J=6.6, 12.4 Hz, 1H), 7.10-7.50 (m,8H), 6.54 (dd, J=6.6, 11.6 Hz, 1H), 4.86 (br, 3H), 3.41-3.67 (m, 4H),2.63-2.92 (m, 3H), 2.02-2.32 (m, 4H), 1.32 (s, 3H), 1.28 (br s, 1H),1.16 (s, 3H), 1.10 (t, J=7.8 Hz, 3H), 1.01 (t, J=7.8 Hz, 3H). MS: (ES)m/z calculated for C₃₃H₃₈F₂N₅O₂ [M+H]⁺ 574.3, found 574.5.

Example 7

Synthesis of1-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

Step a: To a mixture of tert-butyl3,3-dimethyl-4-oxopiperidine-1-carboxylate (4.0 g, 17.6 mmol) in DCM (50mL) at 0° C. was added MgCl₂ (3.4 g, 35.2 mmol),2,5-difluoro-4-nitrobenzoyl chloride (4.3 g, 19.4 mmol) andtriethylamine (4.9 mL, 35.2 mmol) sequentially. The mixture was stirredfor 30 min at 0° C. followed by 1.5 h at room temperature. The mixturewas then cooled to 0° C., quenched with saturated aqueous NH₄Cl andextracted with EtOAc. The organic layer was separated, washed withbrine, dried over Na₂SO₄, and filtered. The solvent was concentratedunder reduced pressure to afford tert-butyl5-(2,5-difluoro-4-nitrobenzoyl)-3,3-dimethyl-4-oxopiperidine-1-carboxylate.MS: (ES) m/z calculated for C₁₉H₂₂F₂N₂O₆Na [M+Na]⁺ 435.1, found 435.1.

Step b: A mixture of tert-butyl5-(2,5-difluoro-4-nitrobenzoyl)-3,3-dimethyl-4-oxopiperidine-1-carboxylate(˜17.6 mmol, crude from step a), (2,6-diethylphenyl)hydrazinehydrochloride (3.5 g, 17.6 mmol) and pyridine (2.1 mL, 26.4 mmol) inMeOH (100 mL) was heated to 45° C. overnight. The mixture was cooled toroom temperature and water was added. The mixture was extracted withEtOAc. The organic layer was separated, washed with brine, dried overNa₂SO₄, and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatography(0 to 60% EtOAc in hexanes) to afford tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₉H₃₅F₂N₄O₄ [M+H]⁺ 541.3, found 541.6.

Step c: A mixture of tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-nitrophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(6.7 g, 6.2 mmol), iron powder (12.0 g, 214.9 mmol) and NH₄Cl (40.0 g,742.1 mmol) in EtOH (100 mL) and water (10 mL) was heated to 90° C. for30 min. The mixture was cooled to room temperature, filtered over Celiteand washed with EtOAc. The filtrate was partitioned between EtOAc andsaturated aqueous NaHCO₃. The organic layer was separated, dried overNa₂SO₄, and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatography(0 to 80% EtOAc in hexanes, then 0 to 30% EtOAc in dichloromethane) toafford tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₉H₃₇F₂N₄O₂ [M+H]⁺ 511.3, found 511.6.

Step d: A mixture of tert-butyl3-(4-amino-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.0 g, 2.0 mmol) and benzoyl isocyanate (0.5 g, 3.4 mmol) in THF (6 mL)was stirred at room temperature for 1 h. The mixture was concentrated ona rotary evaporator under reduced pressure to obtain tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

A mixture of tert-butyl3-(4-(3-benzoylureido)-2,5-difluorophenyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(˜2.0 mmol) from above and K₂CO₃ (1.0 g, 7.2 mmol) in MeOH (20 mL) wasstirred for 2 h at room temperature. The mixture was extracted withIPA:CHCl₃ (1:3). The organic layer was separated, dried over Na₂SO₄, andfiltered. The solvent was concentrated under reduced pressure and theresidue was purified by silica gel flash chromatography (0 to 80% EtOAcin dichloromethane) to afford tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₃₀H₃₈F₂N₅O₃ [M+H]⁺ 554.3, found 554.2

Step e: A mixture of tert-butyl2-(2,6-diethylphenyl)-3-(2,5-difluoro-4-ureidophenyl)-6,6-dimethyl-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(0.6 g, 1.1 mmol) and 4 N HCl in dioxane (30.0 mL, 120.0 mmol) indichloromethane (10 mL) was stirred at room temperature for 1.5 h. Themixture was concentrated under reduced pressure to obtain1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride. MS: (ES) m/z calculated for C₂₅H₃₀F₂N₅O [M+H]⁺ 454.2,found 454.2.

Step f: To a mixture of1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride (100.0 mg, 0.2 mmol), 2,4-bis(trifluoromethyl)benzaldehyde(100.0 mg, 0.8 mmol), and N,N-diisopropylethylamine (0.3 mL, 1.7 mmol)in dichloroethane (10 mL) under magnetic stirring was added NaBH(OAc)₃(250.0 mg, 1.2 mmol) followed by AcOH (two drops). The resulting mixturewas stirred at 45° C. for 3 h. After cooling to room temperature, thereaction mixture was quenched with MeOH, diluted with EtOAc, washed withbrine, and dried over MgSO₄. The solvent was removed under reducedpressure and the residue was purified by preparative TLC (50% EtOAc inhexanes), followed by HPLC (MeCN/H₂O, with 0.1% TFA) to afford1-(4-(5-(2,4-bis(trifluoromethyl)benzyl)-2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluoropheny)urea.¹H NMR (400 MHz, CD₃OD): δ 8.30 (d, J=8.2 Hz, 1H), 7.83-7.95 (m, 3H),7.36 (t, J=7.5 Hz, 1H), 7.20 (d, J=7.7 Hz, 2H), 6.50 (dd, J=6.5, 11.6Hz, 1H), 4.87 (br, 3H), 4.00 (s, 2H), 3.57 (s, 2H), 2.79 (s, 2H),2.22-2.78 (m, 4H), 1.35 (s, 6H), 1.08 (t, J=7.8 Hz, 6H). MS: (ES) m/zcalculated C₃₄H₃₄F₈N₅O [M+H]⁺ 680.3, found 680.6.

Example 8 Synthesis of1-(4-(2-(2,6-diethylphenyl)-5-isobutyryl-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea

A mixture of1-(4-(2-(2,6-diethylphenyl)-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)ureahydrochloride (25 mg, 0.05 mmol), isobutyric acid (80.0 mg, 1.1 mmol),HATU (100 mg, 0.26 mmol) and NEt₃ (0.1 mL, 0.71 mmol) in DMF (1.5 mL)was stirred at 50-70° C. for 40 min. The mixture was cooled to roomtemperature, basified with saturated aqueous NaHCO₃, and extracted withEtOAc. The organic layer was separated, washed with brine, dried overNa₂SO₄, and filtered. The solvent was concentrated under reducedpressure and the residue was purified by silica gel flash chromatography(0 to 100% EtOAc in hexanes) to afford1-(4-(2-(2,6-diethylphenyl)-5-isobutyryl-6,6-dimethyl-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2,5-difluorophenyl)urea.¹H NMR (400 MHz, CDCl₃): δ 8.06 (m, 1H), 7.38 (dd, J=7.6, 7.6 Hz, 1H),7.21 (d, J=8.0 Hz, 2H), 6.55 (m, 1H), 4.58 (s, 2H), 3.30 (m, 3H), 2.95(s, 2H), 2.92 (m, 1H), 2.24 (q, J=7.6 Hz, 4H), 1.58 (s, 6H), 1.04 (m,12H). MS: (ES) m/z calculated for C₂₉H₃₆F₂N₅O₂ [M+H]⁺ 24.3, found 524.6.

Example 9 Synthesis of1-(4-(5-(2-chloro-4-(trifluoromethyl)-phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)urea

Step a: To a stirred solution of tert-butyl4-oxopiperidine-1-carboxylate (5.0 g, 25.1 mmol) in anhydrous THF (30mL) at −78° C. under N2 atmosphere was added 1.0 M LiHMDS solution inTHF (27.5 mL, 27.5 mmol) dropwise. After the solution was stirred for 30min, a solution of 3-fluoro-4-nitrobenzoyl chloride (5.1 g, 25.1 mmol)in THF (6 mL) was added to the mixture. The reaction mixture was stirredat −78° C. for 1 h, and then warmed to room temperature and stirred for2 h. After completion, the reaction mixture was quenched with 1 M NaHSO₄(50 mL), stirred for 10 min, then diluted with EtOAc. The organic layerwas washed with H₂O, brine, dried over Na₂SO₄, and filtered. The solventwas concentrated in vacuo, and the crude material was used directly forthe next step without further purification.

Step b: To a stirred solution of tert-butyl3-(3-fluoro-4-nitrobenzoyl)-4-oxopiperidine-1-carboxylate (6.0 g, 16.4mmol) in EtOH (120 mL) and glacial acetic acid (12.0 mL, 207.9 mmol) wasadded (2-isobutoxy-6- methylphenyl)hydrazine hydrochloride (3.8 g, 16.4mmol) at room temperature. The mixture was stirred for 15 min and thenrefluxed for 3 h. After completion of the reaction, solvent was removedunder reduced pressure and the residue diluted with EtOAc (100 mL). Theorganic layer was washed with aqueous 2 N NaOH, brine, dried overNa₂SO₄, and filtered. The solvent was concentrated under reducedpressure. The residue was purified by silica gel flash chromatography (5to 20% dichloromethane in MeOH) to give tert-butyl3-(3-fluoro-4-nitrophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₈H₃₄FN₄O₅ [M+H]⁻ 525.2, found 525.3

Step c: To a solution of tert-butyl3-(3-fluoro-4-nitrophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(0.8 g, 1.5 mmol) in methanol (20 mL) was added 10% Pd/C (250 mg) atroom temperature. The resulting mixture was stirred under a hydrogen (30psi) atmosphere for 1 h. The reaction mixture was filtered throughCelite and the filtrate was concentrated under reduced pressure to givetert-butyl3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate,which was used directly in the next step without further purification.

Step d: To the solution of tert-butyl3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylatefrom the previous step (0.7 g, 1.4 mmol) in anhydrous THF (10 mL), wasadded K₂CO₃ (0.8 g 5.7 mmol) and acetyl chloride (0.4 g, 5.1 mmol) atroom temperature. The reaction mixture was stirred at room temperaturefor 4 h then diluted with water. The mixture was extracted with EtOAc.The combined organic layers were washed with brine, dried over Na₂SO₄,filtered, and evaporated in vacuo to givetert-butyl-3-(4-acetamido-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.

Step e: To a solution of tert-butyl3-(4-acetamido-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(0.8 g, 1.5 mmol) in dichloromethane (10 mL) was added TFA (0.4 g, 3.6mmol). The resulting mixture was stirred at room temperature for 2 h.After completion of the reaction, the solvent was diluted with water andsaturated aqueous NaHCO₃ and extracted with dichloromethane. The organiclayer was washed with brine and dried over Na₂SO₄. The solvent wasremoved under reduced presure to giveN-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)acetamide.MS: (ES) m/z calculated for C₂₅H₃₀FN₄O₂ [M+H]⁺ 437.2, found 437.3.

Step f: To a mixture ofN-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)acetamide(100.0 mg, 0.25 mmol), 1-bromo-2-chloro-4-(trifluoromethyl)benzene (85.0mg, 0.37 mmol), NaOtBu (47.0 mg, 0.49 mmol) and BINAP (80.0 mg, 0.05mmol) in toluene (3 mL) was added Pd₂(dba)₃ (22 mg, 0.02 mmol). Thereaction mixture was degassed (N₂) for 5 min and stirred under N₂ at105° C. for 6 h. After completion of the reaction, the mixture wascooled to room temperature, diluted with EtOAc, and filtered throughCelite. The filtrate was washed with brine, and dried over Na₂SO₄. Thesolvent was removed under reduced pressure and the residue was purifiedby silica gel flash chromatography (5 to 20% EtOAc in hexanes) to giveN-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)acetamide.MS: (ES) m/z calculated for C₃₂H₃₂ClF₄N₄O₂ [M+H]⁺ 615.2, found 615.3.

Step g: To a solution ofN-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)acetamide(100.0 mg, 0.4 mmol) in MeOH (3 mL) was added 4 N HCl in dioxane (2.5mL, 10.0 mmol). The resulting mixture was stirred at room temperaturefor 2 h. After completion of the reaction, the mixture was diluted withsaturated aqueous NaHCO₃ and extracted with dichloromethane. Thecombined organic layer was washed with brine, and dried over Na₂SO₄. Thesolvent was removed under reduced pressure to give4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluoroaniline.MS: (ES) m/z calculated for C₃₀H₃₀ClF₄N₄O [M+H]⁺ 573.2, found 573.2.

Step h: To a stirred solution of4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluoroaniline(75.0 mg, 0.13 mmol) in anhydrous THF (5 mL) was addedN,N-disopropylethylamine (75.0 mg, 0.65 mmol) and trimethylisocyanate(140.0 mg, 1.2 mmol). The reaction mixture was stirred at roomtemperature for 16 h. After completion, the reaction mixture was dilutedwith EtOAc, washed with brine, and dried over Na₂SO₄. The solvent wasremoved under reduced pressure and the residue was purified by silicagel flash chromatography (20 to 60% EtOAc in hexanes) to afford1-(4-(5-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)-2-fluorophenyl)urea.¹H NMR (400 MHz, CD₃OD): δ 8.00 (t, J=8.6 Hz, 1H), 7.69 (d, J=2.0 Hz,1H), 7.54 (d, J=8.6 Hz, 1H), 7.39 (d, J=8.2 Hz, 1H), 7.33 (t, J=7.8 Hz,1H), 7.0 (d, J=8.2 Hz, 1H), 6.80-6.95 (m, 3H), 4.87 (br, 3H), 4.32 (q,J=9.4 Hz, 2H), 3.65-3.75 (m, 2H), 3.61 (t, J=5.8 Hz, 2H), 2.95-3.10 (m,2H), 2.01 (s, 3H), 1.85-1.98 (m, 1H), 0.86 (d , J=6.6 Hz, 6H). MS: (ES)m/z calculated for C₃₁H₃₁ClF₄N₅O₂ [M+H]⁺ 616.2, found 616.2.

Example 10 Synthesis of1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-5-((2-(trifluoromethyl)phenyl)sulfonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea

Step a: A mixture of tert-butyl3-(4-amino-3-fluorophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(2.4 g, 4.9 mmol), isocyanatotrimethylsilane (7.5 g, 65.6 mmol) andacetic acid (2.87 mL, 47.8 mmol) in dichloromethane (60 mL) was stirredat room temperature for 8 h. The mixture was then basified withsaturated aqueous NaHCO₃ and extracted with dichloromethane. The organiclayer was separated, washed with brine, dried over Na₂SO₄, and filtered.The solvent was concentrated under reduced pressure and the residue waspurified by silica gel flash chromatography (0 to 100% EtOAc in hexanes)to yield tert-butyl3-(3-fluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₉H₃₇FN₅O₄ [M+H]⁺ 538.3, found 538.3.

Step b: A mixture of tert-butyl3-(3-fluoro-4-ureidophenyl)-2-(2-isobutoxy-6-methylphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(1.8 g, 3.3 mmol) and 4 N HCl in dioxane (17.0 mL, 68.0 mmol) indichloromethane (20 mL) was stirred at room temperature for 1 h. Themixture was then concentrated in vacuo to yield1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)ureahydrochloride. MS: (ES) m/z calculated for C₂₄H₂₉FN₅O₂ [M+H]⁺ 438.2,found 438.3

Step c: A mixture of1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)ureahydrochloride (25.0 mg, 0.05 mmol), 2-(trifluoromethyl)benzenesulfonylchloride (30.0 mg, 0.12 mmol) and NEt₃ (0.1 mL, 0.7 mmol) indichloromethane (1.5 mL) was stirred for 30 min at room temperature. Themixture was quenched with water. The obtained mixture was purified bysilica gel flash chromatography (0 to 100% EtOAc in hexanes) to yield1-(2-fluoro-4-(2-(2-isobutoxy-6-methylphenyl)-5-((2-(trifluoromethyl)phenyl)sulfonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenyl)urea.¹H NMR (400 MHz, CDCl₃): δ 8.17 (m, 1H), 7.97 (dd, J=8.4, 8.4 Hz, 1H),7.89 (m, 1H), 7.70 (m, 2H), 7.18 (dd, J=8.0, 8.0 Hz, 1H), 7.07 (d, J=2.8Hz, 1H), 6.68-6.84 (m, 4H), 4.87 (s, 2H), 4.50 (q, J=13.6 Hz, 2H), 3.73(m, 2H), 3.58 (d, J=6.8 Hz, 2H), 2.92 (m, 2H), 1.94 (s, 3H), 1.85 (m,1H), 0.78 (d, J=6.8 Hz, 6H). MS: (ES) m/z calculated forC₃₁H₃₂F₄N₅O₄S[M+H]⁺ 646.2, found 646.2.

Example 11 Synthesis of4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenol

Step a: To a stirred solution of tert-butyl4-oxopiperidine-1-carboxylate (3.0 g, 15.1 mmol) in anhydrous THF (30mL) at −78° C. under N₂ atmosphere was added 1.0 M LiHMDS solution inTHF (16.5 mL, 16.5 mmol) dropwise. After the reaction mixture wasstirred for 30 min, a solution of 4-methoxy benzoylchloride (2.6 g, 15.1mmol) in THF (3 mL) was added to the mixture. The reaction mixture wasstirred at −78° C. for 1 h, and then the mixture was allowed to warm toroom temperature and stirred for 2 h. After completion, the reactionmixture was diluted with EtOH: AcOH (3:1), and (2,6dimethylphenyl)hydrazine hydrochloride (2.6 g, 15.1 mmol) was added. Themixture was stirred for 30 min, and then at 100° C. for 16 h. Aftercompletion of the reaction, the mixture was cooled to room temperatureand diluted with EtOAc. The organic layer was washed with H₂O and thenbrine. The combined organic layer was dried over Na₂SO₄, filtered andconcentrated. The residue was purified by silica gel flashchromatography (5 to 20% EtOAc in hexanes) to givetert-butyl-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate.MS: (ES) m/z calculated for C₂₆H₃₂N₃O₃ [M+H]⁺ 434.2, found 434.3

Step b: To a solution oftert-butyl-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-2,4,6,7-tetrahydro-5H-pyrazolo[4,3-c]pyridine-5-carboxylate(3.5 g, 8.1 mmol) in dichloromethane (30 mL) was added TFA (1.8 g, 16.1mmol). The resulting mixture was stirred at room temperature for 2 h.After completion of the reaction, the solvent was diluted with water andsaturated aqueous NaHCO₃ and extracted with dichloromethane. The organiclayer was washed with brine, and dried over Na₂SO₄. The solvent wasremoved in vacuo to give2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine.MS: (ES) m/z calculated for C₂₁H₂₄N₃O [M+H]⁺ 334.2, found 334.2.

Step c: To a mixeixture of2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine(500.0 mg, 1.5 mmol), 4-tert-butyl-2-bromo-1-methylbenzene (502.0 mg,2.2 mmol), NaOtBu (280.0 mg, 2.9 mmol) and X-Phos (70.0 mg, 2.2 mmol) intoluene (10 mL) was added Pd(OAc)₂ (17.0 mg, 0.07 mmol). The reactionmixture was degassed (N₂) for 5 min and stirred under N2 at 110° C. for16 h. After completion of the reaction, the mixture was cooled to roomtemperature and diluted with EtOAc then filtered through Celite. Thefiltrate was washed with brine, and dried over Na₂SO₄. The solvent wasremoved under reduced pressure and the residue was purified by silicagel flash chromatography (5 to 20% EtOAc in hexanes) to give5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine.¹H NMR (400 MHz, CD₃OD): δ 7.22 (m, 1H), 7.18 (d, J=1.9 Hz, 1H), 7.13(bs, 1H), 7.10 (d, J=2.7 Hz, 1H), 7.09 (s, 1H), 7.05-7.07 (m, 2H),7.0-7.02 (m, 1H), 6.83 (d, J=2.4 Hz, 1H), 6.81 (d, J=2.4 Hz, 1H), 4.13(s, 2H), 3.73 (s, 3H), 3.38 (t, J=5.8 Hz, 2H), 2.89 (t, J=5.8 Hz, 2H),2.29 (s, 3H), 1.98 (s, 6H), 1.26 (s, 9H). MS: (ES) m/z calculated forC₃₂H₃₈N₃O [M+H]⁺ 480.3, found 480.3.

Step d: To a stirred solution of5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-3-(4-methoxyphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine(400.0 mg, 0.8 mmol) in anhydrous dichloromethane (10 mL) at −78° C.under N₂ atmosphere was added 1.0 M BBr₃ solution in dichloromethane(2.1 mL, 2.1 mmol) dropwise. The reaction mixture was stirred at −78° C.for 1 h, and then warmed to room temperature and stirred for 2 h. Aftercompletion, the reaction mixture was quenched with MeOH (2 mL), stirredfor 10 min, then diluted with dichloromethane. The organic layer waswashed with H₂O and brine. The organic layer was dried over Na₂SO₄,filtered and concentrated. The residue was purified by silica gel flashchromatography (5 to 25% EtOAc in hexanes) to give4-(5-(5-(tert-butyl)-2-methylphenyl)-2-(2,6-dimethylphenyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridin-3-yl)phenol.¹H NMR (400 MHz, CD₃OD): δ 7.49 (bs, 1H), 7.25-7.35 (m, 4H), 7.14 (d,J=7.4 Hz, 2H), 6.98 (d, J=2.0 Hz, 1H), 6.96 (d, J=2.0 Hz, 1H), 6.69 (d,J=2.4 Hz, 1H), 6.67 (d, J=2.4 Hz, 1H), 4.69 (bs, 2H), 3.80-3.90 (m, 2H),3.12 (bt, J=7.0 Hz, 2H), 2.44 (s, 3H), 1.99 (s, 6H), 1.30 (s, 9H). MS:(ES) m/z calculated for C₃₁H₃₆N₃O [M+H]⁺ 466.3, found 466.3.

Example 12

This example illustrates the evaluation of the biological activityassociated with specific compounds of the invention.

Materials and Methods

A. Cells 1. C5a Receptor Expressing Cells

a) U937 Cells

U937 cells are a monocytic cell line which express C5aR, and areavailable from ATCC (VA). These cells were cultured as a suspension inRPMI-1640 medium supplemented with 2 mM L-glutamine, 1.5 g/L sodiumbicarbonate, 4.5 g/L glucose, 10 mM HEPES, 1 mM sodium pyruvate, and 10%FBS. Cells were grown under 5% CO₂/95% air, 100% humidity at 37° C. andsubcultured twice weekly at 1:6 (cells were cultured at a density rangeof 1×10⁵ to 2×10⁶ cells/mL) and harvested at 1×10⁶ cells/mL. Prior toassay, cells are treated overnight with 0.5 mM of cyclic AMP (Sigma,Ohio) and washed once prior to use. cAMP treated U937 cells can be usedin C5aR ligand binding and functional assays.

b) Isolated Human Neutrophils

Optionally, human or murine neutrophils can be used to assay forcompound activity. Neutrophils may be isolated from fresh human bloodusing density separation and centrifigation. Briefly, whole blood isincubated with equal parts 3% dextran and allowed to separate for 45minutes. After separation, the top layer is layered on top of 15 mls ofFicoll (15 mls of Ficoll for every 30 mls of blood suspension) andcentrifuged for 30 minutes at 400×g with no brake. The pellet at thebottom of the tube is then isolated and resuspended into PharmLyse RBCLysis Buffer (BD Biosciences, San Jose, Calif.) after which the sampleis again centrifuged for 10 minutes at 400×g with brake. The remainingcell pellet is resuspended as appropriate and consists of isolatedneutrophils.

B. Assays

1. Inhibition of C5aR Ligand Binding

cAMP treated U937 cells expressing C5aR were centrifuged and resuspendedin assay buffer (20 mM HEPES pH 7.1, 140 mM NaCl, 1 mM CaCl₂, 5 mMMgCl₂, and with 0.1% bovine serum albumin) to a concentration of 3×10⁶cells/mL. Binding assays were set up as follows. 0.1 mL of cells wasadded to the assay plates containing 5 μL of the compound, giving afinal concentration of ˜2-10 μM each compound for screening (or part ofa dose response for compound IC₅₀ determinations). Then 0.1 mL of ¹²⁵Ilabeled C5a (obtained from Perkin Elmer Life Sciences, Boston, Mass.)diluted in assay buffer to a final concentration of ˜50 pM, yielding˜30,000 cpm per well, was added, the plates sealed and incubated forapproximately 3 hours at 4° C. on a shaker platform. Reactions wereaspirated onto GF/B glass filters pre-soaked in 0.3% polyethyleneimine(PEI) solution, on a vacuum cell harvester (Packard Instruments;Meriden, Conn.). Scintillation fluid (40 μl; Microscint 20, PackardInstruments) was added to each well, the plates were sealed andradioactivity measured in a Topcount scintillation counter (PackardInstruments). Control wells containing either diluent only (for totalcounts) or excess C5a (1 μg/mL, for non-specific binding) were used tocalculate the percent of total inhibition for compound. The computerprogram Prism from GraphPad, Inc. (San Diego, Calif.) was used tocalculate IC₅₀ values. IC₅₀ values are those concentrations required toreduce the binding of radiolabeled C5a to the receptor by 50%. (Forfurther descriptions of ligand binding and other functional assays, seeDairaghi, et al., J. Biol. Chem. 274:21569-21574 (1999), Penfold, etal., Proc. Natl. Acad. Sci. USA. 96:9839-9844 (1999), and Dairaghi, etal,. J. Biol. Chem. 272:28206-28209 (1997)).

2. Calcium Mobilization

Optionally, compounds may be further assayed for their ability toinhibit calcium flux in cells. To detect the release of intracellularstores of calcium, cells (e.g., cAMP stimulated U937 or neutrophils) areincubated with 3 μM of INDO-1AM dye (Molecular Probes; Eugene, Oreg.) incell media for 45 minutes at room temperature and washed with phosphatebuffered saline (PBS). After INDO-1AM loading, the cells are resuspendedin flux buffer (Hank's balanced salt solution (HBSS) and 1% FBS).Calcium mobilization is measured using a Photon

Technology International spectrophotometer (Photon TechnologyInternational; New Jersey) with excitation at 350 nm and dualsimultaneous recording of fluorescence emission at 400 nm and 490 nm.Relative intracellular calcium levels are expressed as the 400 nm/490 nmemission ratio. Experiments are performed at 37° C. with constant mixingin cuvettes each containing 10⁶ cells in 2 mL of flux buffer. Thechemokine ligands may be used over a range from 1 to 100 nM. Theemission ratio is plotted over time (typically 2-3 minutes). Candidateligand blocking compounds (up to 10 μM) are added at 10 seconds,followed by chemokines at 60 seconds (i.e., C5a; R&D Systems;Minneapolis, Minn.) and control chemokine (i.e., SDF-1α; R&D Systems;Minneapolis, Minn.) at 150 seconds.

3. Chemotaxis Assays

Optionally, compounds may be further assayed for their ability toinhibit chemotaxis in cells. Chemotaxis assays are performed using 5 μmpore polycarbonate, polyvinylpyrrolidone-coated filters in 96-wellchemotaxis chambers (Neuroprobe; Gaithersburg, Md.) using chemotaxisbuffer (Hank's balanced salt solution (HBSS) and 1% FBS). C5aR ligands(i.e., C5a, R&D Systems; Minneapolis, Minn.) are use to evaluatecompound mediated inhibition of C5aR mediated migration. Otherchemokines (i.e., SDF-1≢; R&D Systems; Minneapolis, Minn.) are used asspecificity controls. The lower chamber is loaded with 29 μl ofchemokine (i.e., 0.03 nM C5a) and varying amounts of compound; the topchamber contains 100,000 U937 or neutrophil cells in 20 μl . Thechambers are incubated 1.5 hours at 37° C., and the number of cells inthe lower chamber quantified either by direct cell counts in five highpowered fields per well or by the CyQuant assay (Molecular Probes), afluorescent dye method that measures nucleic acid content andmicroscopic observation.

C. Identification of Inhibitors of C5aR

1. Assay

To evaluate small organic molecules that prevent the C5a receptor frombinding ligand, an assay was employed that detected radioactive ligand(i.e, C5a) binding to cells expressing C5aR on the cell surface (forexample, cAMP stimulated U937 cells or isolated human neutrophils). Forcompounds that inhibited binding, whether competitive or not, fewerradioactive counts are observed when compared to uninhibited controls.

Equal numbers of cells were added to each well in the plate. The cellswere then incubated with radiolabeled C5a. Unbound ligand was removed bywashing the cells, and bound ligand was determined by quantifyingradioactive counts. Cells that were incubated without any organiccompound gave total counts; non-specific binding was determined byincubating the cells with unlabeled ligand and labeled ligand. Percentinhibition was determined by the equation:

% inhibition=(1−[(sample cpm)−(nonspecific cpm)]/[(totalcpm)−(nonspecific cpm)])×100.

2. Dose Response Curves

To ascertain a candidate compound's affinity for C5aR as well as confirmits ability to inhibit ligand binding, inhibitory activity was titeredover a 1×10⁻¹⁰ to 1×10⁻⁴ M range of compound concentrations. In theassay, the amount of compound was varied; while cell number and ligandconcentration were held constant.

D. In Vivo Efficacy Models

The compounds of interest can be evaluated for potential efficacy intreating a C5a mediated conditions by determining the efficacy of thecompound in an animal model. In addition to the models described below,other suitable animal models for studying the compound of interest canbe found in Mizuno, M. et al., Expert Opin. Investig. Drugs (2005),14(7), 807-821, which is incorporated herein by reference in itsentirety.

1. Models of C5a Induced Leukopenia

a) C5a Induced Leukopenia in a Human C5aR Knock-In Mouse Model

To study the efficacy of compounds of the instant invention in an animalmodel, a recombinant mouse can be created using standard techniques,wherein the genetic sequence coding for the mouse C5aR is replaced withsequence coding for the human C5aR, to create a hC5aR-KI mouse. In thismouse, administration of hC5a leads to upregulation of adhesionmolecules on blood vessel walls which bind blood leukocytes,sequestering them from the blood stream. Animals are administered 20ug/kg of hC5a and 1 minute later leukocytes are quantified in peripheralblood by standard techniques. Pretreatment of mice with varying doses ofthe present compounds can almost completely block the hC5a inducedleukopenia.

b) C5a Induced Leukopenia in a Cynomolgus Model

To study the efficacy of compounds of the instant invention in anon-human primate model model, C5a induced leucopenia is studied in acynomolgus model. In this model administration of hC5a leads toupregulation of adhesion molecules on blood vessel walls which bindblood leukocytes, hence sequestering them from the blood stream. Animalsare administered 10 ug/kg of hC5a and 1 minute later leukocytes arequantified in peripheral blood.

Mouse Model of ANCA Induced Vasculitis

On day 0 hC5aR-KI mice are intraveneously injected with 50 mg/kgpurified antibodiy to myeloperoxidase (Xiao et al, J. Clin. Invest. 110:955-963 (2002)). Mice are further dosed with oral daily doses ofcompounds of the invention or vehicle for seven days, then mice aresacrificed and kidneys collected for histological examination. Analysisof kidney sections can show significantly reduced number and severity ofcrescentic and necrotic lesions in the glomeruli when compared tovehicle treated animals.

2. Mouse Model of Choroidal Neovascularization

To study the efficacy of compounds of the instant invention in treatmentof age related macular degeneration (AMD) the bruch membrane in the eyesof hC5aR-KI mice are ruptured by laser photocoagulation (Nozika et al,PNAS 103: 2328-2333 (2006). Mice are treated with vehicle or a dailyoral or appropriate intra-vitreal dose of a compound of the inventionfor one to two weeks. Repair of laser induced damage andneovascularization are assessed by histology and angiography.

3. Rheumatoid Arthritis Models

a) Rabbit Model of Destructive Joint Inflammation

To study the effects of candidate compounds on inhibiting theinflammatory response of rabbits to an intra-articular injection of thebacterial membrane component lipopolysaccharide (LPS), a rabbit model ofdestructive joint inflammation is used. This study design mimics thedestructive joint inflammation seen in arthritis. Intra-articularinjection of LPS causes an acute inflammatory response characterized bythe release of cytokines and chemokines, many of which have beenidentified in rheumatoid arthritic joints. Marked increases inleukocytes occur in synovial fluid and in synovium in response toelevation of these chemotactic mediators. Selective antagonists ofchemokine receptors have shown efficacy in this model (see Podolin, etal., J. Immunol. 169(11):6435-6444 (2002)).

A rabbit LPS study is conducted essentially as described in Podolin, etal. ibid., female New Zealand rabbits (approximately 2 kilograms) aretreated intra-articularly in one knee with LPS (10 ng) together witheither vehicle only (phosphate buffered saline with 1% DMSO) or withaddition of candidate compound (dose 1=50 μM or dose 2=100 μM) in atotal volume of 1.0 mL. Sixteen hours after the LPS injection, knees arelavaged and cells counts are performed. Beneficial effects of treatmentwere determined by histopathologic evaluation of synovial inflammation.Inflammation scores are used for the histopathologic evaluation:1-minimal, 2-mild, 3-moderate, 4-moderate-marked.

b) Evaluation of a Compound in a Rat Model of Collagen Induced Arthritis

A 17 day developing type II collagen arthritis study is conducted toevaluate the effects of a candidate compound on arthritis inducedclinical ankle swelling. Rat collagen arthritis is an experimental modelof polyarthritis that has been widely used for preclinical testing ofnumerous anti-arthritic agents (see Trentham, et al., J. Exp. Med.146(3):857-868 (1977), Bendele, et al., Toxicologic Pathol. 27:134-142(1999), Bendele, et al., Arthritis Rheum. 42:498-506 (1999)). Thehallmarks of this model are reliable onset and progression of robust,easily measurable polyarticular inflammation, marked cartilagedestruction in association with pannus formation and mild to moderatebone resorption and periosteal bone proliferation.

Female Lewis rats (approximately 0.2 kilograms) are anesthetized withisoflurane and injected with Freund's Incomplete Adjuvant containing 2mg/mL bovine type II collagen at the base of the tail and two sites onthe back on days 0 and 6 of this 17 day study. A candidate compound isdosed daily in a sub-cutaneous manner from day 0 till day 17 at aefficacious dose. Caliper measurements of the ankle joint diameter weretaken, and reducing joint swelling is taken as a measure of efficacy.

4. Rat Model of Sepsis

To study the effect of compounds of interest on inhibiting thegeneralized inflammatory response that is associated with a sepsis likedisease, the Cecal Ligation and Puncture (CLP) rat model of sepsis isused. A Rat CLP study is conducted essentially as described in FujimuraN, et al. (American Journal Respiratory Critical Care Medicine 2000;161: 440-446). Briefly described here, Wistar Albino Rats of both sexesweighing between 200-250 g are fasted for twelve hours prior toexperiments. Animals are kept on normal 12 hour light and dark cyclesand fed standard rat chow up until 12 hours prior to experiment. Thenanimals are split into four groups; (i) two sham operation groups and(ii) two CLP groups. Each of these two groups (i.e., (i) and (ii)) issplit into vehicle control group and test compound group. Sepsis isinduced by the CLP method. Under brief anesthesia a midline laparotomyis made using minimal dissection and the cecum is ligated just below theileocaecal valve with 3-0 silk, so the intestinal continuity ismaintained. The antimesinteric surface of the cecum is perforated withan 18 gauge needle at two locations 1 cm apart and the cecum is gentlysqueezed until fecal matter is extruded. The bowel is then returned tothe abdomen and the incision is closed. At the end of the operation, allrats are resuscitated with saline, 3 ml/100 g body weight, givensubcutaneously. Postoperatively, the rats are deprived of food, but havefree access to water for the next 16 hours until they are sacrificed.The sham operated groups are given a laparotomy and the cecum ismanipulated but not ligated or perforated. Beneficial effects oftreatment are measured by histopathological scoring of tissues andorgans as well as measurement of several key indicators of hepaticfunction, renal function, and lipid peroxidation. To test for hepaticfunction aspartate transaminase (AST) and alanine transaminase (ALT) aremeasured. Blood urea nitrogen and creatinine concentrations are studiedto assess renal function. Pro-inflammatory cytokines such as TNF-alphaand IL-lbeta are also assayed by ELISA for serum levels.

5. Mouse SLE Model of Experimental Lupus Nephritis.

To study the effect of compounds of interest on a Systemic LupusErythematosus (SLE), the MRL//pr murine SLE model is used. TheMRL/Mp-Tmfrsf6^(lpr/lpr) strain (MRL/lpr) is a commonly used mouse modelof human SLE. To test compounds efficacy in this model male MRL/lpr miceare equally divided between control and C5aR antagonists groups at 13weeks of age. Then over the next 6 weeks compound or vehicle isadministered to the animals via osmotic pumps to maintain coverage andminimize stress effects on the animals. Serum and urine samples arecollected bi-weekly during the six weeks of disease onset andprogression. In a minority of these mice glomerulosclerosis developsleading to the death of the animal from renal failure. Followingmortality as an indicator of renal failure is one of the measuredcriteria and successful treatment will usually result in a delay in theonset of sudden death among the test groups. In addition, the presenceand magnitude of renal disease may also be monitored continuously withblood urea nitrogen (BUN) and albuminuria measurements. Tissues andorgans were also harvested at 19 weeks and subjected to histopathologyand immunohistochemistry and scored based on tissue damage and cellularinfiltration.

6. Rat Model of COPD

Smoke induced airway inflammation in rodent models may be used to assessefficacy of compounds in Chronic Obstructive Pulmonary Disease (COPD).Selective antagonists of chemokines have shown efficacy in this model(see, Stevenson, et al., Am. J. Physiol Lung Cell Mot Physiol. 288L514-L522, (2005)). An acute rat model of COPD is conducted as describedby Stevenson et al. A compound of interest is administered eithersystemically via oral or IV dosing; or locally with nebulized compound.Male Sprague-Dawley rats (350-400 g) are placed in Perspex chambers andexposed to cigarette smoke drawn in via a pump (50 mL every 30 secondswith fresh air in between). Rats are exposed for a total period of 32minutes. Rats are sacrificed up to 7 days after initial exposure. Anybeneficial effects of treatment are assessed by a decrease inflammatorycell infiltrate, decreases in chemokine and cytokine levels.

In a chronic model, mice or rats are exposed to daily tobacco smokeexposures for up to 12 months. Compound is administered systemically viaonce daily oral dosing, or potentially locally via nebulized compound.In addition to the inflammation observed with the acute model (Stevensenet al.), animals may also exhibit other pathologies similar to that seenin human COPD such as emphysema (as indicated by increased mean linearintercept) as well as altered lung chemistry (see Martorana et al, Am.J. Respir. Crit Care Med. 172(7): 848-53.

7. Mouse EAE Model of Multiple Sclerosis

Experimental autoimmune encephalomyelitis (EAE) is a model of humanmultiple sclerosis. Variations of the model have been published, and arewell known in the field. In a typical protocol, C₅₇BL/6 (Charles RiverLaboratories) mice are used for the EAE model. Mice are immunized with200 ug myelin oligodendrocyte glycoprotein (MOG) 35-55 (PeptideInternational) emulsified in Complete Freund's Adjuvant (CFA) containing4 mg/ml Mycobacterium tuberculosis (Sigma-Aldrich) s.c. on day 0. Inaddition, on day 0 and day 2 animals are given 200 ng of pertussis toxin(Calbiochem) i.v. Clinical scoring is based on a scale of 0-5: 0, nosigns of disease; 1, flaccid tail; 2, hind limb weakness; 3, hind limbparalysis; 4, forelimb weakness or paralysis; 5, moribund. Dosing of thecompounds of interest to be assessed can be initiated on day 0(prophylactic) or day 7 (therapeutic, when histological evidence ofdisease is present but few animals are presenting clinical signs) anddosed once or more per day at concentrations appropriate for theiractivity and pharmacokinetic properties, e.g. 100 mg/kg s.c. Efficacy ofcompounds can be assessed by comparisons of severity (maximum meanclinical score in presence of compound compared to vehicle), or bymeasuring a decrease in the number of macrophages (F4/80 positive)isolated from spinal cords. Spinal cord mononuclear cells can beisolated via discontinuous Percoll-gradient. Cells can be stained usingrat anti-mouse F4/80-PE or rat IgG2b-PE (Caltag Laboratories) andquantitated by FACS analysis using 10 ul of Polybeads per sample(Polysciences).

8. Mouse Model of Kidney Transplantation

Transplantation models can be performed in mice, for instance a model ofallogenic kidney transplant from C₅₇BL/6 to BALB/c mice is described inFaikah Gueler et al, JASN Express, Aug. 27, 2008. Briefly, mice areanesthetized and the left donor kidney attached to a cuff of the aortaand the renal vein with a small caval cuff, and the ureters removed enblock. After left nephrectomy of the recipient, the vascular cuffs areanastomosed to the recipient abdominal aorta and vena cava,respectively, below the level of the native renal vessels. The ureter isdirectly anastomosed into the bladder. Cold ischemia time is 60 min, andwarm ischemia time is 30 min. The right native kidney can be removed atthe time of allograft transplantation or at posttransplantation day 4for long-term survival studies. General physical condition of the miceis monitored for evidence of rejection. Compound treatment of animalscan be started before surgery or immediately after transplantation, egby sub cut injection once daily. Mice are studied for renal function andsurvival. Serum creatinine levels are measured by an automated method(Beckman Analyzer, Krefeld, Germany).

9. Mouse Model of Ischemia/Reperfusion

A mouse model of ischemia/reperfusion injury can be performed asdescribed by Xiufen Zheng et al, Am. J. Pathol, Vol 173:4, October,2008. Briefly, CD1 mice aged 6-8 weeks are anesthetized and placed on aheating pad to maintain warmth during surgery. Following abdominalincisions, renal pedicles are bluntly dissected and a microvascularclamp placed on the left renal pedicle for 25-30 minutes. Followingischemia the clamps are removed along with the right kidney, incisionssutured, and the animals allowed to recover. Blood is collected forserum creatinine and BUN analysis as an indicator of kidney health.Alternatively animal survival is monitored over time. Compound can beadministered to animals before and/or after the surgery and the effectson serum creatinine, BUN or animal survival used as indicators ofcompound efficacy.

10. Mouse Model of Tumor Growth

C₅₇BL/6 mice 6-16 weeks of age are injected subcutaneously with 1×105TC-1 cells (ATCC, VA) in the right or left rear flank. Beginning about 2weeks after cell injection, tumors are measured with calipers every 2-4d until the tumor size required the mice are killed. At the time ofsacrifice animals are subjected to a full necropsy and spleens andtumors removed. Excised tumors are measured and weighed. Compounds maybe administered before and/or after tumor injections, and a delay orinhibition of tumor growth used to assess compound efficacy.

Example 13

The compounds in Table 1, below, were prepared using the methodsdescribed above. Characterization data is provided for each compoundlisted. Activity is provided as follows for the chemotaxis assay usingU937 cells as described herein (Example 12): +, 500 nM<IC_(50;) ++, 50nM<IC₅₀≤500 nM; +++, 5 nM<IC₅₀≤50 nM; and ++++, IC₅₀≤5 nM.

TABLE 1 Structure, Characterization Data and Biological Activity Data ofSpecific Embodiments MS: Mig Compound ES (m/z) IC₅₀ Number Structure[M + H]⁺ (nM) 1.001

550.2 ++++ 1.002

667.2 ++ 1.003

637.2 ++++ 1.004

609.2 +++ 1.005

562.4 +++ 1.006

651.2 +++ 1.007

578.4 ++++ 1.008

564.6 ++++ 1.009

482.6 +++ 1.010

538.4 +++ 1.011

680.6 ++++ 1.012

630.5 ++++ 1.013

585.5 ++++ 1.014

612.4 ++++ 1.015

632.4 ++++ 1.016

666.6 ++++ 1.017

612.4 ++++ 1.018

585.5 ++++ 1.019

524.4 +++ 1.020

510.4 +++ 1.021

632.4 ++++ 1.022

666.6 ++++ 1.023

612.5 ++++ 1.024

612.5 ++++ 1.025

540.6 ++++ 1.026

545.4 +++ 1.027

612.4 ++++ 1.028

602.4 ++++ 1.029

566.6 +++ 1.030

496.5 +++ 1.031

646.6 ++++ 1.032

524.6 ++++ 1.033

652.6 ++++ 1.034

598.6 ++++ 1.035

646.6 ++++ 1.036

598.6 ++++ 1.037

614.6 ++++ 1.038

618.4 ++++ 1.039

602.4 ++++ 1.040

552.5 ++++ 1.041

547.5 + 1.042

664.6 ++++ 1.043

536.5 ++++ 1.044

602.5 ++++ 1.045

614.6 ++++ 1.046

552.6 ++++ 1.047

626.5 ++++ 1.048

609.4 ++++ 1.049

598.6 ++++ 1.050

630.6 ++++ 1.051

516.5 ++++ 1.052

646.6 ++++ 1.053

614.4 ++++ 1.054

572.5 ++++ 1.055

652.6 ++++ 1.056

614.4 ++++ 1.057

618.4 ++++ 1.058

614.4 ++++ 1.059

680.6 ++++ 1.060

624.6 +++ 1.061

590.5 ++++ 1.062

670.6 ++++ 1.063

544.6 ++++ 1.064

598.6 ++++ 1.065

584.6 ++++ 1.066

602.4 ++++ 1.067

584.5 ++++ 1.068

648.6 ++++ 1.069

618.5 ++++ 1.070

584.5 ++++ 1.071

560.5 +++ 1.072

534.5 ++ 1.073

584.5 ++++ 1.074

560.5 ++++ 1.075

588.6 +++ 1.076

618.5 ++++ 1.077

652.5 ++++ 1.078

496.5 ++ 1.079

546.5 +++ 1.080

574.5 ++++ 1.081

498.5 +++ 1.082

480.5 ++ 1.083

498.5 +++ 1.084

544.6 ++++ 1.085

546.5 +++ 1.086

470.5 + 1.087

518.3 ++ 1.088

470.5 + 1.089

533.5 ++++ 1.090

540.5 ++++ 1.091

577.5 ++++ 1.092

516.6 ++++ 1.093

548.5 ++++ 1.094

560.5 ++++ 1.095

508.5 ++++ 1.096

516.6 ++++ 1.097

539.3 ++++ 1.098

564.4 ++++ 1.099

576.2 ++++ 1.100

516.6 ++++ 1.101

552.3 + 1.102

534.5 ++++ 1.103

508.1 ++++ 1.104

567.5 ++ 1.105

567.5 +++ 1.106

526.5 ++++ 1.107

526.5 ++++ 1.108

506.3 + 1.109

492.3 ++ 1.110

510.3 + 1.111

572.3 ++++ 1.112

501.0 ++++ 1.113

551.5 ++++ 1.114

554.3 ++++ 1.115

523.5 ++++ 1.116

557.3 + 1.117

556.6 ++++ 1.118

538.5 ++++ 1.119

542.3 ++++ 1.120

500.2 + 1.121

515.2 + 1.122

560.5 ++++ 1.123

555.3 ++++ 1.124

549.5 ++++ 1.125

558.5 ++ 1.126

542.4 ++ 1.127

509.4 ++++ 1.128

539.4 ++++ 1.129

589.2 ++++ 1.130

605.3 ++++ 1.131

590.2 ++++ 1.132

527.1 ++++ 1.133

517.3 ++ 1.134

538.5 +++ 1.135

559.2 + 1.136

586.2 ++++ 1.137

602.6 ++++ 1.138

572.2 ++++ 1.139

576.2 ++++ 1.140

573.3 + 1.141

568.1 ++++ 1.142

573.2 +++ 1.143

613.0 ++++ 1.144

612.2 ++++ 1.145

574.4 ++++ 1.146

516.4 ++++ 1.147

583.1 ++++ 1.148

574.2 ++++ 1.149

574.2 ++++ 1.150

562.2 ++++ 1.151

530.4 ++++ 1.152

548.2 ++++ 1.153

518.2 +++ 1.154

544.1 ++++ 1.155

510.1 +++ 1.156

574.2 ++++ 1.157

574.2 ++++ 1.158

548.2 ++++ 1.159

552.2 ++++ 1.160

534.2 ++++ 1.161

548.4 ++++ 1.162

544.2 ++++ 1.163

612.2 ++++ 1.164

602.4 ++++ 1.165

535.2 ++++ 1.166

576.2 ++++ 1.167

599.0 ++++ 1.168

581.2 ++++ 1.169

585.1 ++++ 1.170

567.1 ++++ 1.171

588.2 +++ 1.172

591.2 ++++ 1.173

585.1 ++++ 1.174

561.1 ++++ 1.175

568.6 ++++ 1.176

547.0 ++++ 1.177

581.6 ++++ 1.178

551.2 ++++ 1.179

563.0 +++ 1.180

543.1 ++++ 1.181

548.3 ++++ 1.182

567.5 ++++ 1.183

551.0 +++ 1.184

559.4 ++++ 1.185

589.5 ++++ 1.186

562.2 ++++ 1.187

583.0 +++ 1.188

605.1 ++++ 1.189

587.1 ++++ 1.190

581.5 ++++ 1.191

575.2 ++++ 1.192

562.2 ++++ 1.193

548.2 ++ 1.194

600.1 ++++ 1.195

562.2 ++++ 1.196

614.2 ++++ 1.197

594.4 ++++ 1.198

578.3 ++++ 1.199

560.3 ++++ 1.200

544.2 ++++ 1.201

571.1 ++++ 1.202

593.3 ++++ 1.203

623.2 ++++ 1.204

482.2 +++ 1.205

605.2 ++++ 1.206

563.2 ++++ 1.207

574.3 ++++ 1.208

598.5 ++++ 1.209

523.3 ++ 1.210

601.5 ++++ 1.211

615.2 ++++ 1.212

496.3 ++++ 1.213

563.6 ++++ 1.214

526.3 ++++ 1.215

492.3 +++ 1.216

504.3 +++ 1.217

635.2 ++++ 1.218

575.3 ++++ 1.219

478.3 +++ 1.220

573.3 ++++ 1.221

478.3 ++ 1.222

635.2 ++++ 1.223

518.3 ++++ 1.224

547.2 ++++ 1.225

596.2 ++++ 1.226

608.2 + 1.227

553.1 ++++ 1.228

567.2 ++++ 1.229

646.2 +++ 1.230

508.3 +++ 1.231

494.3 +++ 1.232

548.3 ++++ 1.233

532.3 + 1.234

535.3 +++ 1.235

464.2 +++ 1.236

571.2 ++++ 1.237

578.2 +++ 1.238

534.2 +++ 1.239

522.3 ++++ 1.240

597.2 ++++ 1.241

616.2 ++++ 1.242

619.2 ++++ 1.243

490.3 ++++ 1.244

566.2 ++++ 1.245

555.3 +++ 1.246

556.3 +++ 1.247

508.3 ++++ 1.248

522.3 ++ 1.249

551.2 ++++ 1.250

565.1 ++++ 1.251

551.2 +++ 1.252

583.5 ++++ 1.253

478.3 ++++ 1.254

534.3 ++++ 1.255

508.3 ++++ 1.256

534.3 ++++ 1.257

508.3 ++++ 1.258

565.2 +++ 1.259

494.3 +++ 1.260

518.3 + 1.261

606.5 + 1.262

548.2 ++++ 1.263

619.2 ++ 1.264

480.3 ++ 1.265

466.2 ++ 1.266

611.2 + 1.267

613.3 +++ 1.268

597.3 +++ 1.269

617.2 ++++ 1.270

601.2 ++++ 1.271

565.2 +++ 1.272

520.3 ++++ 1.273

494.3 +++ 1.274

508.3 ++++ 1.275

617.3 ++++ 1.276

617.3 ++++ 1.277

664.2 ++++ 1.278

582.2 +++ 1.279

574.3 +++ 1.280

602.3 +++ 1.281

590.3 ++++ 1.282

460.3 +++ 1.283

481.2 + 1.284

572.3 +++ 1.285

514.3 +++ 1.286

576.3 +++ 1.287

539.2 +++ 1.288

601.3 ++++ 1.289

611.3 + 1.290

508.3 +++ 1.291

548.2 +++ 1.292

582.2 +++ 1.293

548.2 +++ 1.294

558.3 +++ 1.295

562.3 +++ 1.296

579.2 ++++ 1.297

582.2 +++ 1.298

628.2 ++ 1.299

630.2 ++++ 1.300

592.3 ++++ 1.301

553.2 +++ 1.302

574.2 +++ 1.303

592.3 +++ 1.304

574.3 +++ 1.305

592.3 +++ 1.306

592.3 ++++ 1.307

564.3 +++ 1.308

542.3 ++++ 1.309

562.3 ++++ 1.310

546.3 +++ 1.311

581.2 +++ 1.312

558.3 ++++ 1.313

544.3 +++ 1.314

592.3 +++ 1.315

612.4 ++++ 1.316

546.2 ++++ 1.317

598.2 ++++ 1.318

582.2 ++++ 1.319

592.3 ++++ 1.320

562.4 ++++ 1.321

578.2 ++++ 1.322

583.2 ++++ 1.323

562.3 ++++ 1.324

578.3 ++++ 1.325

612.2 ++++ 1.326

562.3 ++++ 1.327

612.4 ++++ 1.328

578.2 ++++ 1.329

596.2 ++++ 1.330

592.3 ++++ 1.331

592.3 ++++ 1.332

680.2 ++++ 1.333

646.2 ++++ 1.334

664.2 ++++ 1.335

646.3 ++++ 1.336

488.3 +++ 1.337

644.2 +++ 1.338

578.3 +++ 1.339

633.2 ++++ 1.340

579.2 +++ 1.341

566.2 +++ 1.342

632.2 ++++ 1.343

632.2 ++++ 1.344

598.3 ++++ 1.345

578.3 ++++ 1.346

584.2 +++ 1.347

553.3 ++++ 1.348

553.3 +++ 1.349

501.2 ++ 1.350

562.2 +++ 1.351

596.3 ++++ 1.352

566.2 +++ 1.353

599.2 +++ 1.354

558.5 +++ 1.355

553.3 +++ 1.356

546.3 +++ 1.357

646.3 ++++ 1.358

596.3 ++++ 1.359

558.3 ++++ 1.360

542.3 ++++ 1.361

578.3 ++++ 1.362

612.2 ++++ 1.363

572.4 ++ 1.364

569.5 + 1.365

552.3 ++++ 1.366

570.3 +++ 1.367

578.2 +++ 1.368

582.2 +++ 1.369

582.2 +++ 1.370

553.3 +++ 1.371

524.3 +++ 1.372

542.3 +++ 1.373

544.2 +++ 1.374

583.2 +++ 1.375

598.2 +++ 1.376

538.3 +++ 1.377

578.2 +++ 1.378

582.2 +++ 1.379

598.2 +++ 1.380

522.3 +++ 1.381

504.3 +++ 1.382

510.3 +++ 1.383

569.3 ++ 1.384

592.3 +++ 1.385

476.3 ++ 1.386

583.3 +++ 1.387

564.5 +++ 1.388

536.5 +++ 1.389

492.3 + 1.390

552.3 +++ 1.391

530.2 +++ 1.392

566.3 +++ 1.393

504.3 ++ 1.394

551.3 ++++ 1.395

553.3 +++ 1.396

552.3 +++ 1.397

502.3 ++ 1.398

552.3 +++ 1.399

476.3 ++ 1.400

564.5 + 1.401

550.5 ++++ 1.402

536.5 + 1.403

522.5 ++ 1.404

502.3 +++ 1.405

490.3 +++ 1.406

502.3 ++ 1.407

564.6 +++ 1.408

469.2 ++ 1.409

538.3 +++ 1.410

488.3 ++ 1.411

505.3 + 1.412

564.2 ++++ 1.413

598.2 +++ 1.414

518.3 ++ 1.415

504.3 ++ 1.416

476.3 + 1.417

462.2 ++ 1.418

578.3 +++ 1.419

556.3 +++ 1.420

447.2 + 1.421

570.3 ++++ 1.422

586.6 +++ 1.423

528.3 +++ 1.424

534.3 + 1.425

530.3 + 1.426

502.2 ++ 1.427

570.3 ++++ 1.428

490.3 ++ 1.429

552.3 ++ 1.430

538.3 + 1.431

524.2 ++ 1.432

580.3 ++ 1.433

580.3 ++ 1.434

566.3 ++ 1.435

518.3 ++ 1.436

504.3 ++ 1.437

519.3 + 1.438

516.3 ++ 1.439

520.3 +++ 1.440

578.3 +++ 1.441

580.3 ++++ 1.442

508.3 +++ 1.443

565.3 ++ 1.444

552.3 ++++ 1.445

551.3 +++ 1.446

508.3 + 1.447

497.2 ++ 1.448

570.5 +++ 1.449

552.3 + 1.450

524.3 ++ 1.451

528.2 ++ 1.452

466.3 +++ 1.453

537.3 +++ 1.454

552.3 ++++ 1.455

509.3 +++ 1.456

510.2 +++ 1.457

566.3 ++++ 1.458

523.3 +++ 1.459

467.3 +++ 1.460

524.3 ++++ 1.461

484.2 ++ 1.462

524.3 +++ 1.463

494.3 ++++ 1.464

441.2 ++ 1.465

481.2 +++ 1.466

494.3 ++++ 1.467

452.3 ++ 1.468

451.3 ++ 1.469

481.3 ++ 1.470

465.3 +++ 1.471

451.3 +++ 1.472

480.3 ++ 1.473

452.2 ++ 1.474

538.3 ++++ 1.475

510.2 +++ 1.476

481.2 +++ 1.477

479.3 ++ 1.478

480.2 ++ 1.479

467.2 ++ 1.480

495.5 +++ 1.481

437.3 +++ 1.482

496.4 +++ 1.483

495.4 + 1.484

468.3 ++ 1.485

495.5 ++ 1.486

453.4 ++ 1.487

453.4 + 1.488

524.5 + 1.489

467.4 + 1.490

493.5 + 1.491

425.4 ++

While particular embodiments of this invention are described herein,upon reading the description, variations of the disclosed embodimentsmay become apparent to individuals working in the art, and it isexpected that those skilled artisans may employ such variations asappropriate. Accordingly, it is intended that the invention be practicedotherwise than as specifically described herein, and that the inventionincludes all modifications and equivalents of the subject matter recitedin the claims appended hereto as permitted by applicable law. Moreover,any combination of the above-described elements in all possiblevariations thereof is encompassed by the invention unless otherwiseindicated herein or otherwise clearly contradicted by context.

All publications, patent applications, accession numbers, and otherreferences cited in this specification are herein incorporated byreference as if each individual publication or patent application werespecifically and individually indicated to be incorporated by reference.

1. A compound of Formula (I)

or a pharmaceutically acceptable salt thereof, wherein, ring vertex a isN or C(R^(2c)), ring vertex b is N or C(R^(2d)), and ring vertex e is Nor C(R^(2e)), wherein no more than one of a, b and e is N; X¹ isselected from the group consisting of a bond, C₁₋₈alkylene, C(O),C(O)—C₁₋₄alkylene, and S(O)₂; R¹ is selected from the group consistingof a) 5- to 10-membered heteroaryl having from 1 to 4 heteroatoms asring vertices selected from N, O and S; b) C₆₋₁₀ aryl; c) C₃₋₈cycloalkyl; d) 4- to 8-membered heterocycloalkyl having from 1 to 2heteroatoms as ring vertices selected from N, O and S; and e) C₁₋₈alkyl,C₁₋₈alkoxy, C₁₋₈haloalkyl, —C(O)NR^(1a)R^(1b), and —CO₂R^(1a); whereinR^(1a) and R^(1b) are each independently selected from the groupconsisting of hydrogen, C₁₋₈alkyl, C₆₋₁₀ aryl, and —C₁₋₆alkylene-C₆₋₁₀aryl; wherein the group —X¹—R¹ is optionally substituted with 1 to 5R^(x) substituents; R^(2a) and R^(2e) are each independently selectedfrom the group consisting of hydrogen, C₁₋₆ alkyl, C₁₋₆alkoxy,C₁₋₆haloalkyl,—O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl, —C₁₋₆alkyl-O—C₁₋₆ alkyl,—C₁₋₆alkyl-S—C₁₋₆alkyl, CN, and halogen, and at least one of R_(2a) andR^(2e) is other than hydrogen; R^(2b), R^(2c), and R^(2d) are eachindependently selected from the group consisting of hydrogen, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆haloalkyl, —O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl,—C₁₋₆alkyl-O—C₁₋₆ alkyl, —C₁₋₆alkyl-S—C₁₋₆alkyl, cyano, and halogen;each R³ is independently selected from the group consisting of hydroxyl,C₁₋₄alkyl, C₁₋₄haloalkyl and C₁₋₄hydroxyalkyl, and optionally two R³groups on the same carbon atom are combined to form oxo (═O), andoptionally two R³ groups and the carbon atoms they are attached to forma 3-6 membered ring with 0-2 hetereoatoms as ring members selected fromO, N, and S; R⁴ is independently selected from the group consisting of—X²—OR^(4a), —X²—NR^(4a)R^(4b), —X²—CONR^(4a)R^(4b),—X²—NR_(4a)—C(O)R^(4a), —X²—NR^(4a)—C(O)NR^(4a)R^(4b),—X²—NR^(4a)—C(O)OR^(4b); —X²—NR^(4a)—C(O)—C₁₋₃alkylene-OR^(4a) and—X²—NR^(4a)—C(O)—C₁₋₃alkylene-NR^(4a)R^(4b); wherein each X² isindependently a bond, C(O), C₁₋₄alkylene, C(O)—C₁₋₄alkylene, andC₁₋₄alkylene-C(O), and each R^(4a) and R^(4b) is independently selectedfrom the group consisting of hydrogen, C₁₋₄alkyl, and C₁₋₄haloalkyl;each R⁵ is independently selected from the group consisting ofC₁₋₈alkyl, C₁₋₈alkoxy, C₁₋₈haloalkyl, C₁₋₈haloalkoxy, C₁₋₈hydroxyalkyl,halogen, OH, CN, C(O)R^(5a) and CO₂R^(5a); wherein each R^(5a) isindependently selected from the group consisting of hydrogen, C₁₋₄alkyl, and C₁₋₄haloalkyl; each R^(x) is independently selected from thegroup consisting of halogen, CN, C₁₋₄alkyl, C₁₋₄alkoxy, C₁₋₄haloalkyl,C₁₋₄haloalkoxy, C₁₋₄hydroxy, C₂₋₄alkenyl, C₃₋₆cycloalkyl, CO₂—C₁₋₄alkyl,and CONH₂; the subscript m is 0, 1, 2, 3 or 4; and the subscript n is 0,1, 2 or
 3. 2. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein R⁴ is selected from the group consisting of


3. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R⁴ is selected from the group consisting of


4. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R⁴ is selected from the group consisting of


5. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein X¹ is a bond.
 6. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein X¹ is C(O).
 7. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein X¹ is C₁₋₈alkylene.
 8. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein X¹ isC(O)—C₁₋₄alkylene or S(O)₂.
 9. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R¹ is a 5- to10-membered heteroaryl having from 1 to 4 heteroatoms as ring verticesselected from N, O and S; and wherein the group —X¹—R¹ is optionallysubstituted with 1 to 4 R^(x) substituents.
 10. The compound of claim 9,wherein R¹ is selected from the group consisting of pyrazolyl, pyridyl,pyrimidinyl, imidazolyl, thiazolyl, thiadiazolyl and pyrazinyl; andwherein the group —X¹—R¹ is optionally substituted with 1 to 4 R^(x)substituents.
 11. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is C₆₋₁₀aryl; and wherein the group—X¹—R¹ is optionally substituted with 1 to 4 R^(x) substituents.
 12. Thecompound of claim 11, wherein R¹ is phenyl; and wherein the group —X¹—R¹is optionally substituted with 1 to 4 R^(x) substituents.
 13. Thecompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R¹ is C₃₋₈ cycloalkyl; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents.
 14. The compoundof claim 13, wherein R¹ is selected from the group consisting ofcyclobutyl, cyclopentyl and cyclohexyl; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents.
 15. The compoundof claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ isa 4- to 8-membered heterocycloalkyl having from 1 to 2 heteroatoms asring vertices selected from N, O and S; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents.
 16. The compoundof claim 15, wherein R¹ is selected from the group consisting ofoxetanyl, tetrahydrofuranyl, tetrahydropyranyl and morpholinyl; andwherein the group —X¹—R¹ is optionally substituted with 1 to 4 R^(x)substituents.
 17. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein R¹ is selected from the groupconsisting of C₁₋₈alkyl, C₁₋₈alkoxy, C₁₋₈haloalkyl, —C(O)NR^(1a)R^(1b),and —CO₂R^(1a); wherein R^(1a) and R^(1b) are each independentlyselected from the group consisting of hydrogen, C₁₋₈alkyl, C₆₋₁₀ aryl,and —C₁₋₆ alkylene-C₆₋₁₀ aryl; and wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents.
 18. The compoundof claim 1, or a pharmaceutically acceptable salt thereof, wherein R¹ isselected from the group consisting of phenyl, pyridyl, pyrimidinyl, andpyrazinyl; and wherein the group —X¹—R¹ is optionally substituted with 1to 4 R^(x) substituents.
 19. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein ring vertices a and bare CH; R^(2b) is H; ring vertex e is C(R^(2e)), and R^(2a) and R^(2e)are independently selected from the group consisting of C₁₋₆ alkyl,C₁₋₆alkoxy, C₁₋₆haloalkyl,—O—C₁₋₆haloalkyl, —S—C₁₋₆alkyl,—C₁₋₆alkyl-O—C₁₋₆alkyl, —C₁₋₆ alkyl-S—C₁₋₆alkyl, CN, and halogen. 20.The compound of claim 1, or a pharmaceutically acceptable salt thereof,wherein ring vertices a and b are CH; R^(2b) is H; ring vertex e isC(R^(2e)), and R^(2a) and R^(2e) are independently selected from thegroup consisting of C₁₋₆ alkyl, C₁₋₆alkoxy and halogen.
 21. The compoundof claim 1, or a pharmaceutically acceptable salt thereof, wherein n is0, 1 or 2 and each R⁵, when present, is selected from the groupconsisting of F, Cl, CN, C₁₋₄alkyl and C₁₋₄alkoxy.
 22. The compound ofclaim 1, or a pharmaceutically acceptable salt thereof, wherein n is 0,1 or 2 and each R⁵, when present, is selected from the group consistingof F, Cl, CN, CH₃ and OCH₃.
 23. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein m is 0, 1 or 2 andeach R³, when present, is C₁₋₄alkyl.
 24. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R¹ is selected fromthe group consisting of phenyl or pyridyl, wherein the group —X¹—R¹ isoptionally substituted with 1 to 4 R^(x) substituents; ring vertices aand b are CH; R^(2b) is H; ring vertex e is C(R^(2e)), and R^(2a) andR^(2e) are independently selected from the group consisting ofC₁₋₆alkyl, C₁₋₆alkoxy and halogen; m is 0, 1 or 2 and each R³, whenpresent, is CH₃, R⁴ is selected from the group consisting of

n is 0, 1 or 2 and each R⁵, when present, is selected from the groupconsisting of F, Cl, CN, CH₃ and OCH₃.
 25. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R¹ is selected fromthe group consisting of


26. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein —X¹—R¹ is selected from the group consisting of:


27. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R¹ is selected from the group consisting of:


28. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R¹ is selected from the group consisting of:


29. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein R¹ is selected from the group consisting of:


30. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein

is selected from the group consisting of


31. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein n is
 0. 32. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein n is 2 and the two R³groups are on the same carbon atom and are combined to form oxo (═O).33. The compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein said compound is selected from the group consisting of


34. A pharmaceutical composition comprising a compound of claim 1 or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier.
 35. The pharmaceutical composition of claim 34,formulated for oral, intravenous, transdermal or subcutaneousadministration.
 36. The pharmaceutical composition of claim 34, furthercomprising one or more additional therapeutic agents. 37-38. (canceled)39. A method of treating a human suffering from or susceptible to adisease or disorder involving pathologic activation of C5a receptors,comprising administering to the mammal a therapeutically effectiveamount of a compound of claim
 1. 40. The method of claim 39, wherein thedisease or disorder is an inflammatory disease or disorder, anautoimmune disease, or an oncologic disease or disorder.
 41. The methodof claim 39, wherein the disease or disorder is a cardiovascular orcerebrovascular disorder.
 42. (canceled)
 43. The method of claim 39,wherein the disease or disorder is selected from the group consisting ofneutropenia, neutrophilia, Wegener's granulomatosis, microscopicpolyangiitis, C₃-glomerulopathy, C₃-glomerulonephritis, dense depositdisease, membranoproliferative glomerulonephritis, Kawasaki disease,Hemolytic uremic syndrome, atypical hemolytic uremic syndrome (aHUS),tissue graft rejection, hyperacute rejection of transplanted organs,rheumatoid arthritis, systemic lupus erythematosus, lupus nephritis,lupus glomerulonephritis, vasculitis, ANCA vasculitis, autoimmunehemolytic and thrombocytopenic states, immuno vasculitis, Graft versushost disease, lupus nephropathy, Heyman nephritis, membranous nephritis,glomerulonephritis, IGA nephropathy, hidradenitis suppurativa, andMembranoproliferative glomerulonephritis.
 44. The method of claim 39,wherein the disease or disorder is selected from the group consisting ofmelanoma, lung cancer, lymphoma, sarcoma, carcinoma, fibrosarcoma,liposarcoma, chondrosarcoma, osteogenic sarcoma, angiosarcoma,lymphangiosarcoma, synovioma, mesothelioma, meningioma, leukemia,lymphoma, leiomyosarcoma, rhabdomyosarcoma, squamous cell carcinoma,basal cell carcinoma, adenocarcinoma, papillary carcinoma,cystadenocarcinoma, bronchogenic carcinoma, renal cell carcinoma,hepatocellular carcinoma, transitional cell carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, wilm's tumor, pleomorphic adenoma, livercell papilloma, renal tubular adenoma, cystadenoma, papilloma, adenoma,leiomyoma, rhabdomyoma, hemangioma, lymphangioma, osteoma, chondroma,lipoma and fibroma. 45-47. (canceled)